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  1. Detailansicht für Treffer 41 öffnen
    Figure 1—figure supplement 4. Quantification of ArchT activation-induced pH change in actuator cells. ; (A) Schematic diagram showing the process for calibrating pH. (B) Confocal images showing pHluorin (top) and BFP fluorescence (bottom) of cells perfused with buffers with the indicated pH levels. (C) pH calibration curve for mTagBFP-pHluorinCAAX. The GFP/BFP ratio was fitted with a Hill function (n = 10 cells for each point). (D) Schematic diagram and confocal images showing co-expression of mTagBFP-pHluorinCAAX and ArchT-P2A-mRuby3. The yellow circle indicates the stimulation region. (E) Representative traces of the GFP/BFP ratio and pH change upon a 20 s or 4 s stimulation at 561 nm (three stimuli delivered at 3-min intervals) in cells co-expressing mTagBFP-pHluorinCAAX and ArchT-P2A-mRuby3 (green lines) and in cells expressing only mTagBFP-pHluorinCAAX (gray line). Error bars indicate the s.e.m. from triplicate experiments. (F) Summary of the estimated peak change in pH in the experiments shown in (E) based on the calibration curve shown in (C) (n = 10 cells per group). The scale bars represents10 μm. A, actuator cells; R, receiver cells.
    2019
    unknown
    Wie komme ich dran?
  2. Detailansicht für Treffer 42 öffnen
    Figure 4—figure supplement 2. Characterization of clonal ASNA1 A149V cells. ; (A) Clonal K562 lines containing the mutant A149V (C460T), synonymous A149A (C461T), and wildtype ASNA1 alleles were generated by single cell sorting of edited pools. Homozygous ASNA1 alleles were confirmed by Sanger sequencing. (B) Whole lysates were prepared from clonal wildtype, ASNA1 A149V (C460T), and ASNA1 A149A (C461T) K562 cells either lacking or transduced with the GFP-2A-RFP-SEC61BTMD TA reporter. Samples were subjected to SDS-PAGE and visualized by immunoblotting (IB). Alpha tubulin served as the loading control. (C) Clonal lines were treated with various concentrations of DHQZ36.1 for 5 days and the number of live cells was then counted by cytometry, using FSC/SSC to determine viability. Error bars represent error of the mean from three replicates. (D) The ricin sensitivity and Retro-2 rescue of clonal K562 lines. Cells were pretreated for 24 hr in the indicated Retro-2 concentration before treated with ricin toxin for 24 hr. 72 hr later live cell number was assessed using cytometry and forward/side scatter. Error bars represent error of the mean from three replicates. (E) Clonal wildtype (black circles), ASNA1 A149V (C460T) (red triangles), and ASNA1 A149A (C461T) (blue triangles) K562 cell lines with the GFP-2A-RFP-SEC61BTMD reporter were pre-treated with DHQZ36.1 for 1 hr prior to induction with dox for approximately 18 hr and FACS analysis. Shown are the dose-response curves for the reporter RFP to GFP ratios as relative means (three experiments) to mock-treated wildtype cells. The dose response was modeled using the four-parameter logistic regression to determine the half maximal effective concentration (EC50 ± standard error). Error bars for the means represent the standard error calculated from the four-parameter logistic regression. (F) Clonal wildtype, ASNA1 A149V (C460T), and ASNA1 A149A (C461T) K562 cell lines with the GFP-2A-RFP-SEC61BTMD reporter were pre-treated with the 10 µM DHQZ5 or mock-treated with DMSO for 1 hr prior to induction with dox for approximately 18 hr and FACS analysis. Shown are bar graphs of reporter RFP to GFP ratios with standard deviations derived from three experiments as relative means to mock-treated wildtype cells. (G) ASNA1KO HEK293T cells expressing GFP-2A-RFP-SEC61BTMD were transiently transfected with indicated BFP-ASNA1 variants or BFP. The resulting transfected cells and untransfected wildtype control were pre-treated with 10 µM Retro-2 for 1 hr prior to induction with dox for 24 hr and FACS analysis. Shown are bar graphs of the means ± standard deviation (three experiments) of RFP to GFP ratios normalized to untransfected wildtype.
    2019
    unknown
    Wie komme ich dran?
  3. Detailansicht für Treffer 43 öffnen
    Figure 9. Gain of protein toxicity via autophagy inhibition in prnp-FUS transgenic mice. ; (A) Representative images of lumbar spinal cord sections from hemizygote and transgene homozygote FUSWT transgenic animals stained with p62 (SQSTM1) (green) and HA for human FUS transgene (grey). Scale bar is 20 μm. Cytoplasmic accumulations of p62 were visible in the ventral horn motor neurons of the spinal cord in transgene homozygous mice, but not other genotypes. (B) Schematics of autophagy induction and progression (flux) assay. Double-tagged LC3 (mCherry-GFP-LC3) were used to visualize the induction and progression of autophagy. Upon autophagy induction, LC3 is post-translationally modified with a lipid group and localize to autophagosome. Autophagosomes progress to autolysosomes by fusing with lysosome. Fluorescent signal of GFP is pH-sensitive and is quenched in autolysosome. (C) Representative images of Neuro2A cells co-transfected with BFP (blue fluorescent protein) or BFP-tagged wild type FUS with mCherry-GFP-LC3 (left panel) or immuno-stained with p62 (right panel). Nutrient-limiting medium was used to induce autophagy. (D) Quantification of autophagy induction under nutrient-limiting medium conditions. Over-expression of wild type and ALS-linked mutants (R514G and R521C) in FUS inhibit autophagy induction. (E) Quantification of autophagy flux based on the numbers of autophagosome and autolysosome. Overexpression of FUS inhibits autophagy flux. (F) Quantification of p62 accumulation in cells transfected with BFP or BFP-tagged FUS.
    2019
    unknown
    Wie komme ich dran?
  5. Detailansicht für Treffer 45 öffnen
    Figure 6—figure supplement 1. Membrane traffic performance of two proton-pumps compared with ArchT. ; (A) Confocal images showing the expression of two new proton pumps besides ArchT in HEK293T cells. Proton pumps were fused with BFP at the C-terminus and co-expressed with pHluorinCAAX. (B) The normalized line-scanning plots of the fluorescence signals in both blue and green channels. (C) Relative colocalization were measured by Pearson’s colocalization ratios of the Autuator-BFP according to pHluorinCAAX (n = 28 for each protein). The scale bars represent 10 μm. ***p
    2019
    unknown
    Wie komme ich dran?
  6. Detailansicht für Treffer 46 öffnen
    Figure 7. GPI-anchored protein acutely misfolded at the cell surface is not degraded. ; (A) Cells transiently transfected with FKBP*-YFP-GPI for 48 hr were grown in the presence or absence of 2 µM Shield1, lysed, and immunoblotted using anti-GFP (which also detects YFP). (B) Cells stably expressing inducible FKBP*-YFP-GPI grown with or without Shield1 were incubated on ice with 200 nM Alexa647-conjugated anti-GFP Nb and analyzed by flow cytometry for Nb fluorescence (left) or total YFP fluorescence (right). (C) Schematic of experimental strategy for simultaneousy monitoring the fate of inducibly misfolded FKBP* in the cytosol and on the cell surface. The surface FKBP* is tagged with YFP and the cytosolic FKBP* is tagged with tagBFP. (D) Cells transiently transfected with FKBP*-YFP-GPI and cytosolic FKBP*-BFP for 48 hr were grown in the presence or absence of 2 µM Shield1. Following labeling on ice with Nb-FLAG, cells were washed and incubated for a further two hours at 37°C. During this incubation period, half of the cells previously grown in Shield1 were subjected to washout (w/o) by omitting Shield1 and further supplementing the medium with excess recombinant FKBP* to act as a molecular ‘sink’ to capture any residual Shield1. After the incubation period, half of the cells were treated with extracellular trypsin to digest surface-exposed proteins before inactivating the trypsin. Following lysis, Nb-FLAG and cytosolic FKBP*-BFP were detected by immunoblotting. (E) Cells transiently transfected with FKBP*-YFP-GPI and cytosolic FKBP*-BFP for 48 hr were grown in the presence or absence of 2 µM Shield1. In a subset of cells, Shield1 was withdrawn as in panel D for varying periods of time, after which surface-localized FKBP*-YFP-GPI was stained with Alexa647-Nb. Levels of cytosolic FKBP*-BFP and surface-localized FKBP*-YFP-GPI were monitored via flow cytometry. 2.5 µg/ml Brefeldin A was included in the medium during the time course to prevent export of newly-synthesized protein.
    2019
    unknown
    Wie komme ich dran?
  7. Detailansicht für Treffer 47 öffnen
    Figure 3—figure supplement 2. Cav1.3s is recruited to Kv2-induced EPJs. ; (A) TIRF images of a HEK293T cell cotransfected with the short isoform of Cav1.3 (GFP-Cav1.3 (green), BFP-SEC61β (blue) and auxiliary subunits Cavβ3 and Cavα2δ (not shown). Scalebar is 10 μm and holds for all large panels in figure. Pseudocolored intensity profiles of GFP-Cav1.3 and BFP-SEC61β, from the boxed area in the merged image, are shown to the right of merged image. (scale bar: 2.5 μm and holds for all pseudocolored intensity profiles in figure). (B) TIRF images of HEK293T cells cotransfected with DsRed-Kv2.1 (red), GFP-Cav1.3 (green), BFP-SEC61β (blue) and auxiliary subunits Cavβ3 and Cavα2δ (not shown). Pseudocolored intensity profiles of DsRed-Kv2.1, GFP-Cav1.3 and BFP-SEC61β, from the boxed area in the merged image, are shown to the right of merged image. (C) TIRF images of a HEK293T cell cotransfected with DsRed-Kv2.2 (red), GFP-Cav1.3 (green), BFP-SEC61β (blue) and auxiliary subunits Cavβ3 and Cavα2δ (not shown). Pseudocolored intensity profiles of DsRed-Kv2.2, GFP-Cav1.3 and BFP-SEC61β from the boxed area in the merged image, are shown to the right of merged image. (D) Optical sections of HEK293T cells transfected with and immunolabeled for Cav3.1 alone (upper panel) or with Kv2.1 (lower panels) (scale bar: 10 μm and holds for all panels). (E) Pearson’s correlation coefficient (PCC) values of Cav3.1 and Kv2.1 immunolabeling or Cav1.3-GFP and DsRed-Kv2.1 fluorescence (each point represents a single cell; ****p
    2019
    unknown
    Wie komme ich dran?
  8. Detailansicht für Treffer 48 öffnen
    Figure 3. VPS13A is localized at the ER-mitochondria interface. ; (A) HEK293T cells were co-transfected with VPS13A-Myc and the ER marker GFP-VAP-A. Cells were stained with anti-Myc (red) and DAPI (blue). A’ shows higher magnification of the inserts in A. (B) Representative single stack image of HEK293T cells expressing the ER marker BFP-Sec61B and VPS13A-GFP. Mitochondria were labeled using Mitotracker red. White arrowheads indicate the enrichment of VPS13A at the ER-mitochondria interface. Magenta arrows indicate BFP-Sec61B positive ER tubules, negative for VPS13A-GFP and not in close association with mitochondria. (C) Representative single stack image of HEK293T cells expressing mCherry-VAP-A (ER marker) and VPS13A-GFP. Mitochondria were labeled using TOMM20 antibody. White arrowheads indicate the enrichment of VPS13A at areas positive for ER and mitochondria markers. C’ shows higher magnification of the insert in C. (D) Representative time-lapse images of HEK293T cells expressing VPS13A-GFP and mCherry-VAP-A for 48 hr (Video 2). White arrowheads points to continuous dynamic associations of VPS13A-GFP and mCherry VAP-A. Scale bars = 10 μm (A, C, D), and 2 μm (B).
    2019
    unknown
    Wie komme ich dran?
  9. Detailansicht für Treffer 49 öffnen
    ChrisTrivedi/BFP-Metagenomics-workflow: Manuscript Release
    ChrisTrivedi
    2020
    Online Elektronische Ressource
    Wie komme ich dran?
  10. Detailansicht für Treffer 50 öffnen
    Biomass-Derived Humin-like Furanic Polymers as an Effective UV-Shielding Agent for Optically Transparent Thin-Film Composites
    Rajathsing Kalusulingam (10428764) ; Sampath Gajula (10428767) ; et al.
    2021
    academicJournal
    Wie komme ich dran?
  11. Detailansicht für Treffer 51 öffnen
    Pr-Doping Motivating the Phase Transformation of the BaFeO 3 ‑ δ Perovskite as a High-Performance Solid Oxide Fuel Cell Cathode
    Yunjie Gou (10686988) ; Guangdong Li (653653) ; et al.
    2021
    academicJournal
    Wie komme ich dran?
  12. Detailansicht für Treffer 52 öffnen
    Non-linear momentum transfer of single microtubules and small networks investigated by multiple traps and BFP tracking (Conference Presentation)
    Rohrbach, Alexander ; Koch, Matthias D. ; et al.
    In: Optical Trapping and Optical Micromanipulation XV, 2018
    Konferenz
    Wie komme ich dran?
  13. Detailansicht für Treffer 53 öffnen
    ChrisTrivedi/BFP-Characterization-VSEARCH-workflow: BFP VSEARCH workflow & Zenodo DOI Release
    ChrisTrivedi
    2018
    Online Elektronische Ressource
    Wie komme ich dran?
  14. Detailansicht für Treffer 54 öffnen
    Figure 3. Engagement of integrin αMβ2 (CD11b/CD18) is necessary for formation of the actin cuff. ; After incubation with Candida-BFP hyphae, RAW-Dectin1 cells were fixed and extracellular C. albicans stained using Alexa594-conjugated concanavalin A (red). For panels (A–G) F-actin was stained using fluorescent phalloidin (blue), and actin cuff location indicated with a dashed box or bracket. (A) Anti-CD11b immunostaining (green). Inset: Colocalization of actin cuff with CD11b, in yellow. Scale bar: 5 μm. (B) Anti-CD18 immunostaining (green). Inset: Colocalization of actin cuff with CD18, in yellow. Scale bar: 5 μm. (C) Visualization of transfected Talin-GFP. Inset: Colocalization of actin cuff with talin, in yellow. Scale bar: 10 μm. (D) Immunostaining of endogenous vinculin (green). Inset: Colocalization of actin cuff with vinculin, overlaid in yellow. Scale bar: 10 μm. (E) Immunostaining of endogenous HS1 (green). Scale bar: 10 μm. (F–H) Internalization of Candida-BFP hyphae was allowed to proceed in the presence of the CD11b blocking antibody M1/70 or an isotype-matched (rat IgG2b) control antibody. Following phagocytosis, extracellular C. albicans was stained using Alexa594-conjugated concanavalin A (red), and actin stained using fluorescent phalloidin (blue). Immunostaining (green) for rat IgG2b isotype control (F, left panel) or M1/70 (G, left panel). Scale bars: 5 μm. Images shown are representative of at least 3 experiments of each kind. (H) The number of C. albicans hyphae that were fully internalized or partially internalized with actin cuffs per 37.5x field was counted by confocal microscopy. Average number of C. albicans per field was 11.7 ± 0.5. For each condition, four independent experiments were quantified, with ≥15 fields counted per replicate. p value was calculated using the unpaired, 2-tailed students t-test. Data are means ±SEM.
    2018
    unknown
    Wie komme ich dran?
  15. Detailansicht für Treffer 55 öffnen
    Figure 5. Signaling associated with actin polymerization at the phagocytic cup formed around C. albicans hyphae. ; After incubation with Candida-BFP hyphae, RAW-Dectin1 cells were fixed and extracellular C. albicans stained using fluorescent concanavalin A. (A) Phosphotyrosine (pTyrosine) was detected by immunostaining (green). F-actin was visualized using TdTom-F-Tractin (red); concanavalin A (blue). Inset: Colocalization of actin cuff with pTyrosine, in yellow. Image is representative of ≥30 fields from ≥3 separate experiments. (B) Phospho-SFK (Y418) was detected by immunostaining (green); concanavalin A (red). Inset: Colocalization of actin cuff with pSFK, in yellow. (C) Phospho-PYK2 (Y402) was detected by immunostaining (green); concanavalin A (red). Inset: Colocalization of actin cuff with pPYK2, in yellow. (D) Phospho-FAK (Y397) was detected by immunostaining (green); concanavalin A (red). Inset: Colocalization of actin cuff with pFAK, in yellow. Images in B, C and D are representative of ≥30 fields from ≥2 separate experiments of each type. (E) Effect of tyrosine kinase inhibitors on actin cuff formation. RAW-Dectin1 cells were allowed to adhere Candida-BFP hyphae for 15 min and then incubated 45 min in the presence of vehicle, PP2, piceatannol or PF573228. Following phagocytosis, extracellular C. albicans was stained using concanavalin A, and actin stained with phalloidin. The number of C. albicans hyphae that were fully internalized or partially internalized with actin cuffs per 94.5x field was counted by confocal microscopy. Average number of C. albicans per field was 3.4 ± 0.6. For each condition, three independent experiments were quantified, with ≥4 fields counted per replicate. p value was calculated using unpaired, 2-tailed students t-test. Data are means ±SEM. (F) Active Rac/Cdc42 were visualized using PAK(PBD)-YFP as a probe (green). Actin was stained using fluorescent phalloidin (blue); concanavalin A (red). Inset: Colocalization of actin cuff with PAK(PBD), in yellow. Image is representative of ≥30 fields from ≥3 separate experiments. Scale bars: 10 μm. (G) Effect of actin assembly inhibitors on actin cuff formation. RAW-Dectin1 cells were allowed to adhere Candida-BFP hyphae for 15 min, then incubated 45 min in the presence of vehicle, CK-666 or SMI-FH2. Following phagocytosis, extracellular C. albicans was stained using concanavalin A, and actin stained with phalloidin. The number of C. albicans hyphae that were fully internalized or partially internalized with actin cuffs per 94.5x field was counted by confocal microscopy. Average number of C. albicans per field as in (E). For each condition, three independent experiments were quantified, with ≥5 fields counted per replicate. p value was calculated using the unpaired, 2-tailed students t-test. Data are means ±SEM.
    2018
    unknown
    Wie komme ich dran?
  16. Detailansicht für Treffer 56 öffnen
    Figure 3. Postsynaptic receptors to DA9 maintain their localization and are aligned with DA9 SV clusters after axotomy. ; (A) Labeling of DA9 axon and ACR-12 in GABA motor neurons in intact animals (2d old adults). Scale bars = 10 μm. (B) Labeling of DA9 axon and ACR-12 in GABA motor neurons in axotomized animals. Scale bars = 10 μm. (C) ACR-12::GFP puncta number in the putative DA9 synaptic region (~85 μm anterior from the most posterior ACR-12 puncta in the dorsal nerve cord) in both intact and axotomized animals. ns, not significant. Unpaired t test. (D) Length of posterior DA9 axon without ACR-12 in both intact and axotomized animals. ns, not significant. Unpaired t test. (E) BFP::RAB-3 puncta in DA9 axon appose ACR-12::GFP puncta in the postsynaptic GABA motor neurons in intact animals. Scale bars = 10 μm. (F) BFP::RAB-3 puncta in DA9 axon appose ACR-12::GFP puncta in the postsynaptic GABA motor neurons 48 hr after axotomy. Scale bars = 10 μm. (G) Percentage of BFP::RAB-3 apposing ACR-12::GFP in intact and axotomized animals. ns, not significant. Unpaired t test.
    2018
    unknown
    Wie komme ich dran?
  17. Detailansicht für Treffer 57 öffnen
    Figure 6. Distribution of phosphoinositides and endo-lysosomal markers. ; After incubation with Candida-BFP hyphae, RAW-Dectin1 cells were fixed and extracellular C. albicans stained using Alexa594-conjugated concanavalin A (red). Actin was stained using fluorescent phalloidin (blue), and the location of the actin cuff is indicated by the dashed square. Visualization of: (A) PtdIns(4,5)P2 using PLCδ-PH-GFP; (B) PtdIns(3,4,5)P3/PtdIns(3,4)P2 using AKT-PH-GFP, inset: colocalization of actin cuff with AKT-PH, in yellow; (C) PtdIns(3)P using PX-GFP, inset: colocalization of actin cuff with PX, in yellow; (D) LC3-GFP, inset: colocalization of actin cuff with LC3, in yellow; (E) Rab7-GFP, inset: colocalization of actin cuff with Rab7, in yellow; (F) immunostained LAMP1 (green), inset: colocalization of actin cuff with LAMP1, in yellow. Scale bars: 10 μm. Images are representative of ≥30 fields from ≥3 separate experiments of each type. (G–H) 3D rendering of a RAW-Dectin1 cell with a partially internalized C. albicans hypha. After incubation with Candida-BFP, RAW-Dectin1 cells were fixed and extracellular portions of the hyphae were stained using concanavalin A (blue). (G) C. albicans (white) visualized with actin immunostaining (red). (H) Same 3D rendering as in (G), visualizing LAMP1 immunostaining (green), actin (red) and concanavalin A (blue). Scale bar: 5 μm.
    2018
    unknown
    Wie komme ich dran?
  18. Detailansicht für Treffer 58 öffnen
    Figure 4. PexRD54 accumulates at perihaustorial autophagosomes. ; (A–C) Confocal images of haustoriated plant cells marked by REM1.3. GFP:PexRD54 showed frequent perihaustorial puncta (A) unlike its ATG8 interaction motif mutant PexRD54AIM2 (B), which rarely labelled perihaustorial puncta and showed cytoplasmic distribution similar to GFP:EV (C). (D–F) In haustoriated cells marked by RFP:REM1.3, GFP:ATG8CL labelled autophagosomes fully overlapped with perihaustorial BFP:PexRD54 puncta (D). In contrast, BFP:PexRD54AIM2 mainly remained cytoplasmic and mostly did not show perihaustorial puncta that overlap with ATG8CL autophagosomes (E), similar to BFP:EV control (F). Images shown are maximum projections of 8, 12, 10, 8, 5, and 9 frames with 1 μm steps from top to bottom rows, respectively. White arrowheads point to haustoria. Scale bars, 10 μm. Images are obtained 3–4 dpi. (G) PexRD54/ATG8CL labelled autophagosomes accumulate across the EHM. Single focal plane CLSM images show regions where the EHM and the tonoplast (dotted line) are parted away and the cytosol is no longer a thin layer between the two membranes. Autophagosomes co-labelled by BFP:PexRD54 and GFP:ATG8CL associate with the EHM marked by RFP:REM1.3. Green arrowheads in overlay panel point to BFP:PexRD54 labelled autophagosomes that associate with the EHM marked by RFP:REM1.3. Scale bar, 10 μm.
    2018
    unknown
    Wie komme ich dran?
  20. Detailansicht für Treffer 60 öffnen
    Figure 4. Assessing the contribution of C. albicans cell wall components to actin cuff formation. ; (A) RAW or RAW-Dectin1 cells were incubated with Candida-BFP hyphae that had been either untreated or serum-opsonized. Following phagocytosis, extracellular C. albicans was stained using concanavalin A, and actin stained with phalloidin. The number of C. albicans hyphae that were fully internalized or partially internalized with actin cuffs per 37.5x field was counted by confocal microscopy. Average number of C. albicans per field was 15.7 ± 1.3. For each condition, three independent experiments were quantified, with ≥4 fields counted per replicate. p value was calculated using the unpaired, 2-tailed students t-test. Data are means ±SEM. (B) RAW-Dectin1 cells were allowed to internalize Candida-BFP hyphae in the presence or absence of laminarin or mannan. For laminarin, RAW-Dectin1 cells were also allowed to adhere C. albicans 15 min prior to the addition of laminarin, as indicated. Other details as in A. Average number of C. albicans per field was 12.9 ± 0.7. (C–D) Evaluation of C. albicans GRACE strain cell wall mutants for actin cuff formation. GRACE strains were induced to form hyphae in the absence or presence of doxycycline (DOX) to repress target gene expression, and incubated with RAW-Dectin1 cells. Following phagocytosis, monolayers were fixed and C. albicans stained with 10 μg mL−1 calcofluor white (white), extracellular C. albicans stained using concanavalin A (red), and actin stained with phalloidin (green). Image in C is representative of ≥30 fields from ≥3 separate experiments of each type. Scale bar: 5 μm. (D) The number of C. albicans hyphae that were fully internalized or partially internalized with actin cuffs per 37.5x field was counted by confocal microscopy, and the average number per field calculated. Average number of C. albicans per field was 20.6 ± 0.6. For each condition, three independent experiments were quantified, with ≥4 fields counted per replicate. p value was calculated using the unpaired, 2-tailed students t-test. Data are means ±SEM. (E) Role of C. albicans β-(1,3)-glucan in actin cuff formation. The GRACE wild-type strain was incubated and induced to form hyphae in the presence or absence of 5 ng mL−1 caspofungin and incubated with RAW-Dectin1 cells for phagocytosis. Following phagocytosis, cells were fixed and C. albicans stained with 10 μg mL−1 calcofluor white (white), extracellular C. albicans stained using fluorescent concanavalin A (red), and actin stained with fluorescent phalloidin (green). Image is representative of ≥30 fields from ≥3 separate experiments. Scale bar: 5 μm. (F) The effect of β-(1,3)-glucan synthase inhibition on actin cuff formation. Hyphae were prepared as in (E), with varying concentrations of caspofungin, as indicated. Phagocytosis, fixation and staining as in (E). Other details as in (A). Average number of C. albicans per field was 19.7 ± 0.8. (G) Actin cuffs are observed during phagocytosis of A. fumigatus hyphae. After incubation with hyphae, monolayers were fixed and A. fumigatus stained with 10 μg mL−1 calcofluor white (white). Actin stained with phalloidin (green). Image representative of ≥30 fields from ≥2 separate experiments. Scale bar: 5 μm.
    2018
    unknown
    Wie komme ich dran?
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