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Deletion of Fkh2 in B. bassiana affects cell cycle and septation pattern of multicullular hyphae and transcriptional profiles of related genes. ( A ) Bright and fluorescent images of hyphae stained with the cell-wall-specific dye <t>calcofluor</t> white (upper row) and the nucleic-acid-specific dye DAPI (lower row). Scale bars: 10 μm. ( B , C ) Relative transcript levels (RTL) of cell cycle and septation related genes in the 3-day-old SDB cultures of Fkh2 mutants versus WT, respectively. The asterisked bar in each three-bar group differ significantly from two others unmarked (Tukey’s HSD, P
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1) Product Images from "Transcriptional control of fungal cell cycle and cellular events by Fkh2, a forkhead transcription factor in an insect pathogen"

Article Title: Transcriptional control of fungal cell cycle and cellular events by Fkh2, a forkhead transcription factor in an insect pathogen

Journal: Scientific Reports

doi: 10.1038/srep10108

Deletion of Fkh2 in B. bassiana affects cell cycle and septation pattern of multicullular hyphae and transcriptional profiles of related genes. ( A ) Bright and fluorescent images of hyphae stained with the cell-wall-specific dye calcofluor white (upper row) and the nucleic-acid-specific dye DAPI (lower row). Scale bars: 10 μm. ( B , C ) Relative transcript levels (RTL) of cell cycle and septation related genes in the 3-day-old SDB cultures of Fkh2 mutants versus WT, respectively. The asterisked bar in each three-bar group differ significantly from two others unmarked (Tukey’s HSD, P
Figure Legend Snippet: Deletion of Fkh2 in B. bassiana affects cell cycle and septation pattern of multicullular hyphae and transcriptional profiles of related genes. ( A ) Bright and fluorescent images of hyphae stained with the cell-wall-specific dye calcofluor white (upper row) and the nucleic-acid-specific dye DAPI (lower row). Scale bars: 10 μm. ( B , C ) Relative transcript levels (RTL) of cell cycle and septation related genes in the 3-day-old SDB cultures of Fkh2 mutants versus WT, respectively. The asterisked bar in each three-bar group differ significantly from two others unmarked (Tukey’s HSD, P

Techniques Used: Staining

2) Product Images from "Investigation of Mating Pheromone–Pheromone Receptor Specificity in Lentinula edodes"

Article Title: Investigation of Mating Pheromone–Pheromone Receptor Specificity in Lentinula edodes

Journal: Genes

doi: 10.3390/genes11050506

Effect of PHBs on the monokaryotic S1–10 strain of Lentinula edodes . Expression of the downstream genes in the mating pathway was monitored time-dependently. PHB1 or PHB9 (20 μg/mL) was added to actively growing mycelia of L. edodes S1–10 strain at 25 °C. Inset pictures show the microscopic image of the mushroom mycelia after the treatment of PHB1 or PHB9. The mycelia were stained with calcofluor. The circles indicate clamp connections. Error bars indicate standard deviations of means. The statistical significance of the mean difference is indicated on the bar with asterisks (** for p ≤ 0.01, and **** for p ≤ 0.0001).
Figure Legend Snippet: Effect of PHBs on the monokaryotic S1–10 strain of Lentinula edodes . Expression of the downstream genes in the mating pathway was monitored time-dependently. PHB1 or PHB9 (20 μg/mL) was added to actively growing mycelia of L. edodes S1–10 strain at 25 °C. Inset pictures show the microscopic image of the mushroom mycelia after the treatment of PHB1 or PHB9. The mycelia were stained with calcofluor. The circles indicate clamp connections. Error bars indicate standard deviations of means. The statistical significance of the mean difference is indicated on the bar with asterisks (** for p ≤ 0.01, and **** for p ≤ 0.0001).

Techniques Used: Expressing, Staining

3) Product Images from "Bacterial GtfB Augments Candida albicansAccumulation in Cross-Kingdom Biofilms"

Article Title: Bacterial GtfB Augments Candida albicansAccumulation in Cross-Kingdom Biofilms

Journal: Journal of Dental Research

doi: 10.1177/0022034517714414

Microscopic comparison of haploid and diploid Candida albicans biofilms in the presence and absence of Streptococcus mutans. C. albicans in the single-and mixed-species biofilms were fixed and stained with calcofluor white (in blue), and the S. mutans in mixed-species biofilms were stained with propidium iodide (in red) for confocal laser scanning microscopy (CLSM) visualization (images I-IV and IX-XII). Images V-VIII and XIII-XVI represent only the C. albicans cells in the single-or mixed-species biofilms. ( A ) The presence of S. mutans notably enhanced the biofilm formation of bcr1Δ (IV and VIII) and bcr1Δ/Δ (XII and XVI) mutants in comparison to the absence of S. mutans (II, VI and X, XIV). Haploid and diploid parent strains, GZY803 (III and VII) and SC5314 (XI and XV), also showed a slight increase in biofilm formation in the presence of S. mutans . ( B ) The biovolumes of C. albicans single-and mixed-species biofilms were compared using the Imaris software.
Figure Legend Snippet: Microscopic comparison of haploid and diploid Candida albicans biofilms in the presence and absence of Streptococcus mutans. C. albicans in the single-and mixed-species biofilms were fixed and stained with calcofluor white (in blue), and the S. mutans in mixed-species biofilms were stained with propidium iodide (in red) for confocal laser scanning microscopy (CLSM) visualization (images I-IV and IX-XII). Images V-VIII and XIII-XVI represent only the C. albicans cells in the single-or mixed-species biofilms. ( A ) The presence of S. mutans notably enhanced the biofilm formation of bcr1Δ (IV and VIII) and bcr1Δ/Δ (XII and XVI) mutants in comparison to the absence of S. mutans (II, VI and X, XIV). Haploid and diploid parent strains, GZY803 (III and VII) and SC5314 (XI and XV), also showed a slight increase in biofilm formation in the presence of S. mutans . ( B ) The biovolumes of C. albicans single-and mixed-species biofilms were compared using the Imaris software.

Techniques Used: Staining, Confocal Laser Scanning Microscopy, Software

Glucosyltransferase B (GtfB) increases biofilm formation in Candida albicans. The biofilm formation in the presence of GtfB was evaluated using ( A ) visual observation, ( B ) XTT reduction assay, ( C ) colony-forming unit (CFU) counting, and ( D ) confocal microscopy. Confocal laser scanning microscopy (CLSM) images show C. albicans cells stained with Calcofluor white (in blue) and GtfB-derived glucans labeled with Alexa Fluor 647–labeled dextran conjugate (in red). Adding cell-free, purified GtfB significantly increased biofilm formation in C. albicans . This figure is available in color online.
Figure Legend Snippet: Glucosyltransferase B (GtfB) increases biofilm formation in Candida albicans. The biofilm formation in the presence of GtfB was evaluated using ( A ) visual observation, ( B ) XTT reduction assay, ( C ) colony-forming unit (CFU) counting, and ( D ) confocal microscopy. Confocal laser scanning microscopy (CLSM) images show C. albicans cells stained with Calcofluor white (in blue) and GtfB-derived glucans labeled with Alexa Fluor 647–labeled dextran conjugate (in red). Adding cell-free, purified GtfB significantly increased biofilm formation in C. albicans . This figure is available in color online.

Techniques Used: Confocal Microscopy, Confocal Laser Scanning Microscopy, Staining, Derivative Assay, Labeling, Purification

4) Product Images from "Mitochondria reorganization upon proliferation arrest predicts individual yeast cell fate"

Article Title: Mitochondria reorganization upon proliferation arrest predicts individual yeast cell fate

Journal: eLife

doi: 10.7554/eLife.35685

The capacity to enter quiescence does not correlate with a cell’s replicative age or density. ( A–D ) WT cells expressing Ilv3-RFP were grown 7 d in YPD, stained with calcofluor white and refed onto a YPD microscope pad. ( A ) Percentage of mother or daughter cells with vesicular mitochondria able to form a bud within 6 hr after refeeding; n > 270, N = 3. Representative image series are shown. Number in yellow indicate mothers, in blue, daughters. ( B ) Distribution of the different mother cell categories according to the number of bud scars within the population of mother cells; n > 600, N = 3. ( C ) Percentage of cells with vesicular (green) or globular (red) mitochondrial network within each mother cell category; n > 400, N = 3. ( D ) Individual cell’s ability to form a new bud within 6 hr after refeeding onto a YPD microscope pad according to the mother cell replicative age category; n > 400, N = 3. ( E ) and ( F ) WT cells expressing Ilv3-RFP were grown 7 d in YPD then centrifuged through a percoll density gradient. ( E ) The percentage of dead cells and cells with globular mitochondria were scored; n > 500, N = 2. ( F ) A non-fractionated population (unFract.) was stained with ConA-AlexaFluor647 (pink). Within the fractionated population, the lower fraction (lower F.) was stained with ConA-FITC (green). These two cell subpopulations were re-mixed with the upper fraction subpopulation (upper F.) that was left unstained (blue). Cells were then stained with calcofluor white and allowed to re-proliferate on a YPD microscope pad. The percentage of individual cells with vesicular mitochondria from each fraction that were able to bud within 6 hr after refeeding was scored (N = 3). A representative image series is shown. On the left a field merging the FITC (green), the AlexaFluor647 (pink) and the calcofluor white (blue) channels is shown. Cells are surrounded by dashed line with the color code indicated above. Bar are 2 μm. Error bars are SD.
Figure Legend Snippet: The capacity to enter quiescence does not correlate with a cell’s replicative age or density. ( A–D ) WT cells expressing Ilv3-RFP were grown 7 d in YPD, stained with calcofluor white and refed onto a YPD microscope pad. ( A ) Percentage of mother or daughter cells with vesicular mitochondria able to form a bud within 6 hr after refeeding; n > 270, N = 3. Representative image series are shown. Number in yellow indicate mothers, in blue, daughters. ( B ) Distribution of the different mother cell categories according to the number of bud scars within the population of mother cells; n > 600, N = 3. ( C ) Percentage of cells with vesicular (green) or globular (red) mitochondrial network within each mother cell category; n > 400, N = 3. ( D ) Individual cell’s ability to form a new bud within 6 hr after refeeding onto a YPD microscope pad according to the mother cell replicative age category; n > 400, N = 3. ( E ) and ( F ) WT cells expressing Ilv3-RFP were grown 7 d in YPD then centrifuged through a percoll density gradient. ( E ) The percentage of dead cells and cells with globular mitochondria were scored; n > 500, N = 2. ( F ) A non-fractionated population (unFract.) was stained with ConA-AlexaFluor647 (pink). Within the fractionated population, the lower fraction (lower F.) was stained with ConA-FITC (green). These two cell subpopulations were re-mixed with the upper fraction subpopulation (upper F.) that was left unstained (blue). Cells were then stained with calcofluor white and allowed to re-proliferate on a YPD microscope pad. The percentage of individual cells with vesicular mitochondria from each fraction that were able to bud within 6 hr after refeeding was scored (N = 3). A representative image series is shown. On the left a field merging the FITC (green), the AlexaFluor647 (pink) and the calcofluor white (blue) channels is shown. Cells are surrounded by dashed line with the color code indicated above. Bar are 2 μm. Error bars are SD.

Techniques Used: Expressing, Staining, Microscopy

Mitochondrial network reorganization in non-proliferating cells. ( A ) Mitochondrial network organization in WT cells expressing Ilv3-RFP as a function of time in YPD (OD 600nm : circles). Cells with a tubular (violet), a fragmented (blue), a vesicular (green), a globular (red) mitochondrial network or no Ilv3-RFP signal (grey) were scored. Representative cells of each category are shown; n > 500, N = 2. ( B ) WT, dnm1Δ , fis1Δ , caf4Δ , mdv1Δ , and atg32Δ cells expressing Ilv3-RFP were grown 7 d in YPD. Histograms display the percentage of Ilv3-RFP positive cells with a vesicular (green) or a globular (red) mitochondrial network; n > 200, N = 2. ( C ) WT cells expressing Ilv3-RFP were grown 7 d in YPD and stained with DAPI. Yellow and white arrows point to stained or unstained mitochondria respectively. ( D ) WT cells grown 7 d in YPD co-expressing Ilv3-RFP and the mitochondrial DNA-binding protein Abf2-GFP or the MICOS complex protein Fcj1-GFP. ( E ) WT cells grown 7 d in YPD co-expressing Ilv3-RFP and the respiratory chain component Cox4-GFP or co-expressing Ilv3-GFP and the ATP synthase subunit Atp14-RFP. ( F ) WT cells expressing Ilv3-RFP were grown in YPD to OD 600nm ~ 1. Cells were then shifted to YPD (black) or YPGE (grey). Cells with globular mitochondria were scored; n > 300, N = 2. ( G ) WT cells expressing Ilv3-RFP were grown in YPD to OD 600nm ~ 1. Cells were then shifted to YPGE. After 4 hr, cells were pulse stained with ConA-FITC and shifted to YPD or YPGE. The percentage of cells with globular mitochondria was scored within both ConA-FITC stained (green) and unstained (grey) cell populations; n > 250, N = 2. ( H ) Cells from ( G ) were stained with calcofluor white concomitantly to the ConA-FITC pulse or 15 hr after the transfer to YPGE. The percentage of ConA-FITC positive mother (orange), unbudded daughter (dark blue) and budding daughter (light blue) cells were scored; n > 250, N = 2. Histograms show means, error bars are SD, bar is 2 μm.
Figure Legend Snippet: Mitochondrial network reorganization in non-proliferating cells. ( A ) Mitochondrial network organization in WT cells expressing Ilv3-RFP as a function of time in YPD (OD 600nm : circles). Cells with a tubular (violet), a fragmented (blue), a vesicular (green), a globular (red) mitochondrial network or no Ilv3-RFP signal (grey) were scored. Representative cells of each category are shown; n > 500, N = 2. ( B ) WT, dnm1Δ , fis1Δ , caf4Δ , mdv1Δ , and atg32Δ cells expressing Ilv3-RFP were grown 7 d in YPD. Histograms display the percentage of Ilv3-RFP positive cells with a vesicular (green) or a globular (red) mitochondrial network; n > 200, N = 2. ( C ) WT cells expressing Ilv3-RFP were grown 7 d in YPD and stained with DAPI. Yellow and white arrows point to stained or unstained mitochondria respectively. ( D ) WT cells grown 7 d in YPD co-expressing Ilv3-RFP and the mitochondrial DNA-binding protein Abf2-GFP or the MICOS complex protein Fcj1-GFP. ( E ) WT cells grown 7 d in YPD co-expressing Ilv3-RFP and the respiratory chain component Cox4-GFP or co-expressing Ilv3-GFP and the ATP synthase subunit Atp14-RFP. ( F ) WT cells expressing Ilv3-RFP were grown in YPD to OD 600nm ~ 1. Cells were then shifted to YPD (black) or YPGE (grey). Cells with globular mitochondria were scored; n > 300, N = 2. ( G ) WT cells expressing Ilv3-RFP were grown in YPD to OD 600nm ~ 1. Cells were then shifted to YPGE. After 4 hr, cells were pulse stained with ConA-FITC and shifted to YPD or YPGE. The percentage of cells with globular mitochondria was scored within both ConA-FITC stained (green) and unstained (grey) cell populations; n > 250, N = 2. ( H ) Cells from ( G ) were stained with calcofluor white concomitantly to the ConA-FITC pulse or 15 hr after the transfer to YPGE. The percentage of ConA-FITC positive mother (orange), unbudded daughter (dark blue) and budding daughter (light blue) cells were scored; n > 250, N = 2. Histograms show means, error bars are SD, bar is 2 μm.

Techniques Used: Expressing, Staining, Binding Assay

5) Product Images from "Abnormal Ergosterol Biosynthesis Activates Transcriptional Responses to Antifungal Azoles"

Article Title: Abnormal Ergosterol Biosynthesis Activates Transcriptional Responses to Antifungal Azoles

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2018.00009

erg11 is an essential gene in N. crassa . (A) Schematic diagram of the process of P tcu-1 :: erg11 (NCU02624, encoding sterol 14α-demethylase) strain construction. (B,C) erg11 is required for fungal growth. Conidia of N. crassa bd Ku70 RIP strain (the parental strain), P tcu-1 :: erg11 mutant and the complemented P erg11 :: erg11 mutant were inoculated on Vogel’s plates (B) and Vogel’s slants (C) supplemented with or without CuSO 4 or Cu 2+ chelator BCS and grown at 28°C for 7 days (B) or 30 h (C) , respectively. (D) erg11 inactivation resulted in sterol content changes in N. crassa . After grown for 13.5 h in liquid Vogel’s medium amended with BCS, the mycelium was then transferred to liquid Vogel’s medium amended with BCS and CuSO 4 and treated for 22 h. The sterols were then extracted and subjected to HPLC-MS analysis. The abundance of sterols was calculated on the basis of the chromatogram peak area and normalized using an internal control and sample weight. The results presented here are means of two biological replicates. (E) Conidia of bd Ku70 RIP and P tcu-1 :: erg11 were germinated in Vogel’s minimal medium supplemented with BCS or CuSO 4 for 10 h. Hyphae were imaged after fixing and staining with calcofluor white and DAPI. Clustered nuclei are indicated by arrows and enlarged (below panel on the right).
Figure Legend Snippet: erg11 is an essential gene in N. crassa . (A) Schematic diagram of the process of P tcu-1 :: erg11 (NCU02624, encoding sterol 14α-demethylase) strain construction. (B,C) erg11 is required for fungal growth. Conidia of N. crassa bd Ku70 RIP strain (the parental strain), P tcu-1 :: erg11 mutant and the complemented P erg11 :: erg11 mutant were inoculated on Vogel’s plates (B) and Vogel’s slants (C) supplemented with or without CuSO 4 or Cu 2+ chelator BCS and grown at 28°C for 7 days (B) or 30 h (C) , respectively. (D) erg11 inactivation resulted in sterol content changes in N. crassa . After grown for 13.5 h in liquid Vogel’s medium amended with BCS, the mycelium was then transferred to liquid Vogel’s medium amended with BCS and CuSO 4 and treated for 22 h. The sterols were then extracted and subjected to HPLC-MS analysis. The abundance of sterols was calculated on the basis of the chromatogram peak area and normalized using an internal control and sample weight. The results presented here are means of two biological replicates. (E) Conidia of bd Ku70 RIP and P tcu-1 :: erg11 were germinated in Vogel’s minimal medium supplemented with BCS or CuSO 4 for 10 h. Hyphae were imaged after fixing and staining with calcofluor white and DAPI. Clustered nuclei are indicated by arrows and enlarged (below panel on the right).

Techniques Used: Mutagenesis, High Performance Liquid Chromatography, Mass Spectrometry, Staining

6) Product Images from "Transcriptome analysis of genes involved in secondary cell wall biosynthesis in developing internodes of Miscanthus lutarioriparius"

Article Title: Transcriptome analysis of genes involved in secondary cell wall biosynthesis in developing internodes of Miscanthus lutarioriparius

Journal: Scientific Reports

doi: 10.1038/s41598-017-08690-8

Immunolabeling of crystalline cellulose in an elongating M. lutarioriparius internode. The crystalline cellulose epitope in three sections of M. lutarioriparius internode was immunolabeled with CBM3a. The sections were counterstained with Calcofluor. ( A ) upper internode (UI), ( B ) middle internode (MI), ( C ) basal internode (BI). Bar = 100 μm.
Figure Legend Snippet: Immunolabeling of crystalline cellulose in an elongating M. lutarioriparius internode. The crystalline cellulose epitope in three sections of M. lutarioriparius internode was immunolabeled with CBM3a. The sections were counterstained with Calcofluor. ( A ) upper internode (UI), ( B ) middle internode (MI), ( C ) basal internode (BI). Bar = 100 μm.

Techniques Used: Immunolabeling

7) Product Images from "Protozoan predation drives adaptive divergence in Pseudomonas fluorescens SBW25; ecology meets experimental evolution"

Article Title: Protozoan predation drives adaptive divergence in Pseudomonas fluorescens SBW25; ecology meets experimental evolution

Journal: bioRxiv

doi: 10.1101/2021.07.12.452127

Bacteria evolved under predation develop biofilm morphologies that are comparable to adaptive divergence phenotypes. A) Colony morphologies on LBA (Scale size: 10 mm). B) Visualisation of cellulose production grown on KB agar with a combination of calcofluor staining and microscopy. Scale bar: 30μm. C) Phase contrast image of colony segments shown in figure B. D) Microcosms after three days of growth in KB. E) Graph showing the summed mass of the beads that were added before the biofilm mat broke for each mutant type of interest (N=4, *****P
Figure Legend Snippet: Bacteria evolved under predation develop biofilm morphologies that are comparable to adaptive divergence phenotypes. A) Colony morphologies on LBA (Scale size: 10 mm). B) Visualisation of cellulose production grown on KB agar with a combination of calcofluor staining and microscopy. Scale bar: 30μm. C) Phase contrast image of colony segments shown in figure B. D) Microcosms after three days of growth in KB. E) Graph showing the summed mass of the beads that were added before the biofilm mat broke for each mutant type of interest (N=4, *****P

Techniques Used: Staining, Microscopy, Mutagenesis

8) Product Images from "Biofilm Formation by the Fungal Pathogen Candida albicans: Development, Architecture, and Drug Resistance"

Article Title: Biofilm Formation by the Fungal Pathogen Candida albicans: Development, Architecture, and Drug Resistance

Journal: Journal of Bacteriology

doi: 10.1128/JB.183.18.5385-5394.2001

CSLM images of Calcofluor-stained mature C. albicans biofilms formed on silicone elastomer surface. (a and b) Projection ( xz or side view) of 3-D reconstructed images showing an approximately 450-μm-thick biofilm with a basal layer (10 to 12 μm thick) consisting of yeast cells and a top layer consisting of hyphal elements (arrow). The extracellular material (ECM) is stained with ConA, resulting in the green color. (c and d) Orthogonal images of the basal (c) and upper layers (d). The ECM-derived haziness seen in mature biofilm (e) is absent when the extracellular material is removed (f). Magnification, ×20.
Figure Legend Snippet: CSLM images of Calcofluor-stained mature C. albicans biofilms formed on silicone elastomer surface. (a and b) Projection ( xz or side view) of 3-D reconstructed images showing an approximately 450-μm-thick biofilm with a basal layer (10 to 12 μm thick) consisting of yeast cells and a top layer consisting of hyphal elements (arrow). The extracellular material (ECM) is stained with ConA, resulting in the green color. (c and d) Orthogonal images of the basal (c) and upper layers (d). The ECM-derived haziness seen in mature biofilm (e) is absent when the extracellular material is removed (f). Magnification, ×20.

Techniques Used: Staining, Derivative Assay

9) Product Images from "The role of microsporidian polar tube protein 4 (PTP4) in host cell infection"

Article Title: The role of microsporidian polar tube protein 4 (PTP4) in host cell infection

Journal: PLoS Pathogens

doi: 10.1371/journal.ppat.1006341

Effect of the manipulation of host cell TfR1 expression on E . hellem infection. (A) Immunoblot of TRVb cells (lane 1) and TRVb-1 cells (lane 2) using MAb-TfR1. (B) The CHO cell lines TRVb (a TfR1 knockout) and TRVb-1 (that expresses human TfR1) were infected with E . hellem and infection rates determined as described in Materials and Methods. A significant decrease in infection was observed in TRVb-1 cells compared to TRVb cells, which is consistent with binding to TfR1 being involved in the infection process. (C ) Fluorescent microscopy (using a triple band D/F/TR XF467 Omega Optical filter cube and a 40X objective; insert is a 60X objective) of E . hellem invasion assay at 6 hours post infection. E . hellem infected CHO cell lines, TRVb or TRVb-1, were incubated for 6 hours with spores labeled with Calcofluor white, the cells were then fixed and in situ hybridization with an E . hellem rRNA probe labeled with Alexa Fluor 594 was performed to distinguish intracellular forms (red fluorescence at white arrows) from extracellular adherent spores (blue spores) as described in Materials in Methods. The white arrowheads point to spores, that we believe, are in the process of invading where the sporoplasm is at the edge of the spore, but where intracellular invasion has not yet been completed or occurred. Bars, 10μm. (D) Percent of CHO cell lines TRVb or TRVb-1 infected at 6 hours. A significant decrease (p
Figure Legend Snippet: Effect of the manipulation of host cell TfR1 expression on E . hellem infection. (A) Immunoblot of TRVb cells (lane 1) and TRVb-1 cells (lane 2) using MAb-TfR1. (B) The CHO cell lines TRVb (a TfR1 knockout) and TRVb-1 (that expresses human TfR1) were infected with E . hellem and infection rates determined as described in Materials and Methods. A significant decrease in infection was observed in TRVb-1 cells compared to TRVb cells, which is consistent with binding to TfR1 being involved in the infection process. (C ) Fluorescent microscopy (using a triple band D/F/TR XF467 Omega Optical filter cube and a 40X objective; insert is a 60X objective) of E . hellem invasion assay at 6 hours post infection. E . hellem infected CHO cell lines, TRVb or TRVb-1, were incubated for 6 hours with spores labeled with Calcofluor white, the cells were then fixed and in situ hybridization with an E . hellem rRNA probe labeled with Alexa Fluor 594 was performed to distinguish intracellular forms (red fluorescence at white arrows) from extracellular adherent spores (blue spores) as described in Materials in Methods. The white arrowheads point to spores, that we believe, are in the process of invading where the sporoplasm is at the edge of the spore, but where intracellular invasion has not yet been completed or occurred. Bars, 10μm. (D) Percent of CHO cell lines TRVb or TRVb-1 infected at 6 hours. A significant decrease (p

Techniques Used: Expressing, Infection, Knock-Out, Binding Assay, Microscopy, Invasion Assay, Incubation, Labeling, In Situ Hybridization, Fluorescence

10) Product Images from "Conidial Dihydroxynaphthalene Melanin of the Human Pathogenic Fungus Aspergillus fumigatus Interferes with the Host Endocytosis Pathway"

Article Title: Conidial Dihydroxynaphthalene Melanin of the Human Pathogenic Fungus Aspergillus fumigatus Interferes with the Host Endocytosis Pathway

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2011.00096

Effect of bafilomycin A1 . Lysosomes were loaded with Oregon Green (OG) 488-Dextran. Colocalization of calcofluor white (CFW)-stained conidia (blue) with OG488 (green) showing fusion of phagosomes with lysosomes. Acidification of phagolysosomes was monitored by LysoTracker (red). Images show representative microscopic pictures. Size bar, 2 μm.
Figure Legend Snippet: Effect of bafilomycin A1 . Lysosomes were loaded with Oregon Green (OG) 488-Dextran. Colocalization of calcofluor white (CFW)-stained conidia (blue) with OG488 (green) showing fusion of phagosomes with lysosomes. Acidification of phagolysosomes was monitored by LysoTracker (red). Images show representative microscopic pictures. Size bar, 2 μm.

Techniques Used: Staining

Monitoring of phagolysosomal fusion, acidification of conidia-containing phagosomes, and of intraphagosomal pH . (A) Lysosomes of MH-S and THP-1 cells were chased with Oregon Green (OG) 488-Dextran. Colocalization of calcofluor white (CFW)-stained conidia (blue) with OG488 (green) showing fusion of phagosomes with lysosomes. Acidification of phagolysosomes was studied by LysoTracker (red). Images show representative microscopic pictures. Size bar, 5 μm. (B) Quantification of intracellular conidia residing in mature phagolysosomes of MH-S cells. Data represent the mean + SD from three experiments. (C) Determination of the intraphagosomal pH of conidia-containing phagosomes of MH-S cells. MH-S cells were allowed to phagocytose FITC-labeled conidia. Intracellular pH was calculated by linear regression analysis based on the fluorescence intensity ratio at Ex 495 nm /Ex 450 nm in comparison to a pH standard curve. Values represent the mean + SD of three experiments. * P
Figure Legend Snippet: Monitoring of phagolysosomal fusion, acidification of conidia-containing phagosomes, and of intraphagosomal pH . (A) Lysosomes of MH-S and THP-1 cells were chased with Oregon Green (OG) 488-Dextran. Colocalization of calcofluor white (CFW)-stained conidia (blue) with OG488 (green) showing fusion of phagosomes with lysosomes. Acidification of phagolysosomes was studied by LysoTracker (red). Images show representative microscopic pictures. Size bar, 5 μm. (B) Quantification of intracellular conidia residing in mature phagolysosomes of MH-S cells. Data represent the mean + SD from three experiments. (C) Determination of the intraphagosomal pH of conidia-containing phagosomes of MH-S cells. MH-S cells were allowed to phagocytose FITC-labeled conidia. Intracellular pH was calculated by linear regression analysis based on the fluorescence intensity ratio at Ex 495 nm /Ex 450 nm in comparison to a pH standard curve. Values represent the mean + SD of three experiments. * P

Techniques Used: Staining, Labeling, Fluorescence

Detection of conidia in acidified compartments after phagocytosis by MH-S and THP-1 cells, and neutrophil granulocytes . (A) Percentages of conidia present in acidic phagolysosomes. Calcofluor white (CFW)-labeled conidia were intracellularly colocalized with LysoTracker Red-DND99. Data represent the mean values and SD of three experiments. (B) Representative micrographs showing colocalization of conidia stained using CFW (blue) with acidified compartments visualized by LysoTracker labeling (red). Size bar, 2 μm.
Figure Legend Snippet: Detection of conidia in acidified compartments after phagocytosis by MH-S and THP-1 cells, and neutrophil granulocytes . (A) Percentages of conidia present in acidic phagolysosomes. Calcofluor white (CFW)-labeled conidia were intracellularly colocalized with LysoTracker Red-DND99. Data represent the mean values and SD of three experiments. (B) Representative micrographs showing colocalization of conidia stained using CFW (blue) with acidified compartments visualized by LysoTracker labeling (red). Size bar, 2 μm.

Techniques Used: Labeling, Staining

11) Product Images from "Radiation Tolerance of Pseudanabaena catenata, a Cyanobacterium Relevant to the First Generation Magnox Storage Pond"

Article Title: Radiation Tolerance of Pseudanabaena catenata, a Cyanobacterium Relevant to the First Generation Magnox Storage Pond

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2020.00515

Light microscopy of P. catenata filaments at day 16: A–D were washed twice with normal saline [0.9 g L – 1 NaCl] (A) auto-fluorescence of control culture; (B) calcofluor white stained control culture; (C) auto-fluorescence of irradiated culture; (D) calcofluor white stained irradiated culture; (E) calcofluor white stained unwashed control culture; and (F) calcofluor white stained unwashed irradiated culture. Scale bar denotes 10 μm.
Figure Legend Snippet: Light microscopy of P. catenata filaments at day 16: A–D were washed twice with normal saline [0.9 g L – 1 NaCl] (A) auto-fluorescence of control culture; (B) calcofluor white stained control culture; (C) auto-fluorescence of irradiated culture; (D) calcofluor white stained irradiated culture; (E) calcofluor white stained unwashed control culture; and (F) calcofluor white stained unwashed irradiated culture. Scale bar denotes 10 μm.

Techniques Used: Light Microscopy, Fluorescence, Staining, Irradiation

12) Product Images from "Microglia and amyloid precursor protein coordinate control of transient Candida cerebritis with memory deficits"

Article Title: Microglia and amyloid precursor protein coordinate control of transient Candida cerebritis with memory deficits

Journal: Nature Communications

doi: 10.1038/s41467-018-07991-4

Brain recovery of Candida albicans after intravenous injection. a Wild-type C57BL/6 mice were challenged once intravenously with the indicated doses of C. albicans and the mean colony forming units (CFUs) cultured from whole brain 4 days later were determined. b Clearance of C. albicans from brains of mice after i.v. challenge with 25,000 viable cells over 10 days. c , d Immunofluorescent staining of mouse brain 4 days post i.v. challenge. c Low-magnification view of whole brain after staining with DAPI, IBA1, and GFAP (scale bar: 750 μm) and d PAS staining on a consequent slide demonstrated granuloma-like structures (scale bar: 20 μm). e – i Higher magnification views of the boxed area in c stained with DAPI, IBA1, GFAP, and calcofluor white (CW), respectively. The merged image is shown in h . (Scale bar: 20 μm. n = 4 in b ). Data are representative of three ( a , b ) and six ( c – i ) independent experiments
Figure Legend Snippet: Brain recovery of Candida albicans after intravenous injection. a Wild-type C57BL/6 mice were challenged once intravenously with the indicated doses of C. albicans and the mean colony forming units (CFUs) cultured from whole brain 4 days later were determined. b Clearance of C. albicans from brains of mice after i.v. challenge with 25,000 viable cells over 10 days. c , d Immunofluorescent staining of mouse brain 4 days post i.v. challenge. c Low-magnification view of whole brain after staining with DAPI, IBA1, and GFAP (scale bar: 750 μm) and d PAS staining on a consequent slide demonstrated granuloma-like structures (scale bar: 20 μm). e – i Higher magnification views of the boxed area in c stained with DAPI, IBA1, GFAP, and calcofluor white (CW), respectively. The merged image is shown in h . (Scale bar: 20 μm. n = 4 in b ). Data are representative of three ( a , b ) and six ( c – i ) independent experiments

Techniques Used: Injection, Mouse Assay, Cell Culture, Staining

Amyloid β binds directly to Candida albicans in vivo. a Wild-type mice were challenged with C. albicans as in Fig. 2 , and immunofluorescence staining for DAPI, IBA1, Aβ, and GFAP was performed on brain sections containing fungal granulomas. b Repeat staining of fungal granulomas for DAPI, IBA1, Aβ, and calcofluor white (CW) comparing wild-type to app −/− mice. c , d Aβ 1–42 and 1–40 levels from whole-brain homogenates as assessed by ELISA. ( n = 4, mean ± S.E.M, * p
Figure Legend Snippet: Amyloid β binds directly to Candida albicans in vivo. a Wild-type mice were challenged with C. albicans as in Fig. 2 , and immunofluorescence staining for DAPI, IBA1, Aβ, and GFAP was performed on brain sections containing fungal granulomas. b Repeat staining of fungal granulomas for DAPI, IBA1, Aβ, and calcofluor white (CW) comparing wild-type to app −/− mice. c , d Aβ 1–42 and 1–40 levels from whole-brain homogenates as assessed by ELISA. ( n = 4, mean ± S.E.M, * p

Techniques Used: In Vivo, Mouse Assay, Immunofluorescence, Staining, Enzyme-linked Immunosorbent Assay

C. albicans cerebritis induces upregulation of amyloid precursor protein (APP). Mice were challenged with C. albicans as in Fig. 2 and RNA and protein extracted from whole brain. a qRT-PCR was performed to quantify mRNA for APP over 14 days. b Western blot analysis of APP over the same time period. c Densitometric analysis of the western blot data. d Immunofluorescence images of brain from C. albicans -infected wild-type mice 4 days post i.v. challenge with C. albicans showing staining for DAPI, IBA1, APP, NeuN, and merged images. Images are centered on fungal granulomas similar to those shown in Fig. 2 . e Repeat staining of fungal granulomas for DAPI, IBA1, APP, and calcofluor white (CW) comparing wild-type to app − / − mice. ( n = 4, mean ± S.E.M, * p
Figure Legend Snippet: C. albicans cerebritis induces upregulation of amyloid precursor protein (APP). Mice were challenged with C. albicans as in Fig. 2 and RNA and protein extracted from whole brain. a qRT-PCR was performed to quantify mRNA for APP over 14 days. b Western blot analysis of APP over the same time period. c Densitometric analysis of the western blot data. d Immunofluorescence images of brain from C. albicans -infected wild-type mice 4 days post i.v. challenge with C. albicans showing staining for DAPI, IBA1, APP, NeuN, and merged images. Images are centered on fungal granulomas similar to those shown in Fig. 2 . e Repeat staining of fungal granulomas for DAPI, IBA1, APP, and calcofluor white (CW) comparing wild-type to app − / − mice. ( n = 4, mean ± S.E.M, * p

Techniques Used: Mouse Assay, Quantitative RT-PCR, Western Blot, Immunofluorescence, Infection, Staining

13) Product Images from "Mucosal immunity–mediated modulation of the gut microbiome by oral delivery of probiotics into Peyer’s patches"

Article Title: Mucosal immunity–mediated modulation of the gut microbiome by oral delivery of probiotics into Peyer’s patches

Journal: Science Advances

doi: 10.1126/sciadv.abf0677

Preparation and characterization of EcN@YM. ( A ) LSCM images of EcN@YM. The red channel shows EcN expressing mCherry, and the blue channel indicates YMs stained with calcofluor-white. Scale bars, 20 μm. ( B ) Typical TEM images of EcN and EcN@YM. Scale bars, 2 μm. ( C ) Zeta potentials of EcN, YMs, and EcN@YM, respectively. ** P
Figure Legend Snippet: Preparation and characterization of EcN@YM. ( A ) LSCM images of EcN@YM. The red channel shows EcN expressing mCherry, and the blue channel indicates YMs stained with calcofluor-white. Scale bars, 20 μm. ( B ) Typical TEM images of EcN and EcN@YM. Scale bars, 2 μm. ( C ) Zeta potentials of EcN, YMs, and EcN@YM, respectively. ** P

Techniques Used: Expressing, Staining, Transmission Electron Microscopy

14) Product Images from "Eisosomes promote the ability of Sur7 to regulate plasma membrane organization in Candida albicans"

Article Title: Eisosomes promote the ability of Sur7 to regulate plasma membrane organization in Candida albicans

Journal: Molecular Biology of the Cell

doi: 10.1091/mbc.E16-01-0065

The Pkh2 and Pkh3 kinases prevent formation of elongated eisosomes. (A) Localization of Lsp1-GFP in wild-type control and pkh2∆/∆ pkh3∆/PKH3 cells. The full diploid genotype is listed for this strain to designate that it lacks PKH2 and is heterozygous for PKH3 . Cells were analyzed by fluorescence microscopy, and representative midsections are shown. Right, an enlarged midsection for the pkh2∆/∆ pkh3∆/PKH3 strain, revealing intracellular filaments of Lsp1-GFP. (B) Top view of cells, showing the pattern of Lsp1-GFP in the indicated wild-type or pkh mutant strain. Deletion of the PKH genes resulted in formation of elongated patches of Lsp1-GFP. (C) Cell walls were detected by staining with calcofluor white. (D) Dilutions of the indicated cells were spotted onto agar plates to test the effect on growth of different inhibitory compounds. These data revealed a different pattern of sensitivity to these agents for the pil1∆ lsp1∆ mutant and the pkh2∆/∆ pkh3∆/PKH3 mutant. The top row of assays was carried out with rich YPD medium. The bottom row of spot assays was carried out with SD medium (synthetic medium with dextrose). (E) Plasma membrane furrows were analyzed in the indicated strains by freeze-etching methods and TEM. Strains used for A and B were LSP1-GFP (YLD76-1), LSP1-GFP pkh2∆ (YLD173-2) , LSP1-GFP pkh3∆ (YLD178-1), and LSP1-GFP pkh2∆/∆ pkh3∆/PKH3 (YLD174-4). Strains used for C–E were wild-type control (DIC185), sur7∆ (YJA11), pil1∆ lsp1∆ (YHXW21-1), pkh2∆/∆ pkh3∆PKH3 (YLD146-6-54-1), and pkh2∆/∆ pkh3∆PKH3 plus PKH2 (YLD203-3-3). Bars, 5 μm (A, C), 1 μm (B), 250 nm (E).
Figure Legend Snippet: The Pkh2 and Pkh3 kinases prevent formation of elongated eisosomes. (A) Localization of Lsp1-GFP in wild-type control and pkh2∆/∆ pkh3∆/PKH3 cells. The full diploid genotype is listed for this strain to designate that it lacks PKH2 and is heterozygous for PKH3 . Cells were analyzed by fluorescence microscopy, and representative midsections are shown. Right, an enlarged midsection for the pkh2∆/∆ pkh3∆/PKH3 strain, revealing intracellular filaments of Lsp1-GFP. (B) Top view of cells, showing the pattern of Lsp1-GFP in the indicated wild-type or pkh mutant strain. Deletion of the PKH genes resulted in formation of elongated patches of Lsp1-GFP. (C) Cell walls were detected by staining with calcofluor white. (D) Dilutions of the indicated cells were spotted onto agar plates to test the effect on growth of different inhibitory compounds. These data revealed a different pattern of sensitivity to these agents for the pil1∆ lsp1∆ mutant and the pkh2∆/∆ pkh3∆/PKH3 mutant. The top row of assays was carried out with rich YPD medium. The bottom row of spot assays was carried out with SD medium (synthetic medium with dextrose). (E) Plasma membrane furrows were analyzed in the indicated strains by freeze-etching methods and TEM. Strains used for A and B were LSP1-GFP (YLD76-1), LSP1-GFP pkh2∆ (YLD173-2) , LSP1-GFP pkh3∆ (YLD178-1), and LSP1-GFP pkh2∆/∆ pkh3∆/PKH3 (YLD174-4). Strains used for C–E were wild-type control (DIC185), sur7∆ (YJA11), pil1∆ lsp1∆ (YHXW21-1), pkh2∆/∆ pkh3∆PKH3 (YLD146-6-54-1), and pkh2∆/∆ pkh3∆PKH3 plus PKH2 (YLD203-3-3). Bars, 5 μm (A, C), 1 μm (B), 250 nm (E).

Techniques Used: Fluorescence, Microscopy, Mutagenesis, Staining, Transmission Electron Microscopy

The C. albicans orthologue of S. cerevisiae Slm1/2 localizes to eisosomes but is not needed for proper cell wall morphogenesis. (A) Orf19.3505-GFP (Slm2-GFP) colocalized with Lsp1-RFP, indicating that it is present in eisosomes, consistent with it being the C. albicans orthologue of S. cerevisiae Slm1 and Slm2 proteins. In contrast, Orf19.4043-GFP displayed a diffuse cytoplasmic localization, consistent with it being the orthologue of the S. cerevisiae Ask10 and Rgc1 proteins. (B) In the sur7∆ mutant, Orf19.3035-GFP (Slm2-GFP) localized to punctate clusters, although there were fewer patches than in a wild-type strain. In contrast, Orf19.3035-GFP showed a very different abnormal localization in the eisosome mutant pil1∆ lsp1∆ . The predominant pattern was two major spots at opposite poles of the cells. (C) Left, cells lacking or overexpressing ORF19.3035 ( SLM2 ) do not display cell wall or morphogenesis defects. Cells were stained with calcofluor white. Right, the cells could be induced to form hyphae by treatment with 15% serum for 1.5 h. Strains used were ORF19.3035-GFP LSP1-RFP (YHXW32-1), ORF19.4043-GFP LSP1-RFP (YHXW31-1), ORF19.3505-GFP sur7∆ (YHXW33-1), ORF19.3505 pil1lsp1 (YHXW34-1), orf19.3505∆ (YHXW36-1), and ORF19.3505 overexpressed (YHXW37-1). Bars, 5 μm.
Figure Legend Snippet: The C. albicans orthologue of S. cerevisiae Slm1/2 localizes to eisosomes but is not needed for proper cell wall morphogenesis. (A) Orf19.3505-GFP (Slm2-GFP) colocalized with Lsp1-RFP, indicating that it is present in eisosomes, consistent with it being the C. albicans orthologue of S. cerevisiae Slm1 and Slm2 proteins. In contrast, Orf19.4043-GFP displayed a diffuse cytoplasmic localization, consistent with it being the orthologue of the S. cerevisiae Ask10 and Rgc1 proteins. (B) In the sur7∆ mutant, Orf19.3035-GFP (Slm2-GFP) localized to punctate clusters, although there were fewer patches than in a wild-type strain. In contrast, Orf19.3035-GFP showed a very different abnormal localization in the eisosome mutant pil1∆ lsp1∆ . The predominant pattern was two major spots at opposite poles of the cells. (C) Left, cells lacking or overexpressing ORF19.3035 ( SLM2 ) do not display cell wall or morphogenesis defects. Cells were stained with calcofluor white. Right, the cells could be induced to form hyphae by treatment with 15% serum for 1.5 h. Strains used were ORF19.3035-GFP LSP1-RFP (YHXW32-1), ORF19.4043-GFP LSP1-RFP (YHXW31-1), ORF19.3505-GFP sur7∆ (YHXW33-1), ORF19.3505 pil1lsp1 (YHXW34-1), orf19.3505∆ (YHXW36-1), and ORF19.3505 overexpressed (YHXW37-1). Bars, 5 μm.

Techniques Used: Mutagenesis, Staining

Overexpression of SUR7 partially suppresses the cell wall phenotypes of pil1∆ lsp1∆ cells. Wild type, pil1∆ lsp1∆ , and versions of pil1∆ lsp1∆ carrying one extra copy of SUR7 integrated into the genome (+1x SUR7 ) or two extra copies of SUR7 (+2x SUR7 ) were analyzed. (A) Spot assays showing the growth of the strains indicated on the left to the compounds listed above. YPD indicates the rich nutrient medium, which was used as the base medium for all of the assays. (B) Wild-type and indicated mutant cells were stained with calcofluor white, and then cell wall structures were photographed by fluorescence microscopy. (C) Relative proportions of cells with cell wall invaginations detected by calcofluor white staining as described in Figure 7B . (D) Relative sensitivity to fluconazole for the indicated strains. Cells were spread on an agar plate, and then filter disks containing 0.25, 0.5, or 1 μg fluconazole were placed on top and the plates incubated for 2 d at 37°C. Strains used were wild-type control (DIC185), pil1∆ lsp1∆ (YHXW21-1), pil1∆ lsp1∆ +1xSUR7 (YHXW45-1), and pil1∆ lsp1∆ +2xSUR7 (YHXW46-1). Bar, 5 μm.
Figure Legend Snippet: Overexpression of SUR7 partially suppresses the cell wall phenotypes of pil1∆ lsp1∆ cells. Wild type, pil1∆ lsp1∆ , and versions of pil1∆ lsp1∆ carrying one extra copy of SUR7 integrated into the genome (+1x SUR7 ) or two extra copies of SUR7 (+2x SUR7 ) were analyzed. (A) Spot assays showing the growth of the strains indicated on the left to the compounds listed above. YPD indicates the rich nutrient medium, which was used as the base medium for all of the assays. (B) Wild-type and indicated mutant cells were stained with calcofluor white, and then cell wall structures were photographed by fluorescence microscopy. (C) Relative proportions of cells with cell wall invaginations detected by calcofluor white staining as described in Figure 7B . (D) Relative sensitivity to fluconazole for the indicated strains. Cells were spread on an agar plate, and then filter disks containing 0.25, 0.5, or 1 μg fluconazole were placed on top and the plates incubated for 2 d at 37°C. Strains used were wild-type control (DIC185), pil1∆ lsp1∆ (YHXW21-1), pil1∆ lsp1∆ +1xSUR7 (YHXW45-1), and pil1∆ lsp1∆ +2xSUR7 (YHXW46-1). Bar, 5 μm.

Techniques Used: Over Expression, Mutagenesis, Staining, Fluorescence, Microscopy, Incubation

The pil1∆ lsp1∆ mutant forms large invaginations of the cell wall. (A) The cells were stained with calcofluor white, and then cell wall structures were photographed by fluorescence microscopy. Note that the pil1∆ and lsp1∆ mutants displayed very little altered cell wall, whereas the pil1∆ lsp1∆ double mutant displayed cell wall invaginations comparable to those of the sur7∆ mutant. White bar, 5 μm. (B) Cell sections detected by transmission electron microscopy. Note the presence of thicker cell walls and invaginations of cell wall in both the pil1∆ lsp1∆ double mutant and the sur7∆ mutant. Black arrows on the outer edges of the cells point to regions where there are spiky invaginations in the sur7∆ mutant and the large, round cell wall invagination in the pil1∆ lsp1∆ mutant. The white arrows point to the cell wall tubes. Black bar, 1 μm. The strains used were wild type (DIC185), lsp1∆ (YLD77-11-24-1), pil1∆ (YLD73-9-2-1), pil1∆ lsp1∆ (YHXW21-1), sur7∆ (YJA11), pil1∆ lsp1∆ with a copy of LSP1 reintegrated (YHXW23-1), pil1∆ lsp1∆ with a copy of PIL1 integrated (YHXW22-1), and seg1∆ (YLD182-40-14-3).
Figure Legend Snippet: The pil1∆ lsp1∆ mutant forms large invaginations of the cell wall. (A) The cells were stained with calcofluor white, and then cell wall structures were photographed by fluorescence microscopy. Note that the pil1∆ and lsp1∆ mutants displayed very little altered cell wall, whereas the pil1∆ lsp1∆ double mutant displayed cell wall invaginations comparable to those of the sur7∆ mutant. White bar, 5 μm. (B) Cell sections detected by transmission electron microscopy. Note the presence of thicker cell walls and invaginations of cell wall in both the pil1∆ lsp1∆ double mutant and the sur7∆ mutant. Black arrows on the outer edges of the cells point to regions where there are spiky invaginations in the sur7∆ mutant and the large, round cell wall invagination in the pil1∆ lsp1∆ mutant. The white arrows point to the cell wall tubes. Black bar, 1 μm. The strains used were wild type (DIC185), lsp1∆ (YLD77-11-24-1), pil1∆ (YLD73-9-2-1), pil1∆ lsp1∆ (YHXW21-1), sur7∆ (YJA11), pil1∆ lsp1∆ with a copy of LSP1 reintegrated (YHXW23-1), pil1∆ lsp1∆ with a copy of PIL1 integrated (YHXW22-1), and seg1∆ (YLD182-40-14-3).

Techniques Used: Mutagenesis, Staining, Fluorescence, Microscopy, Transmission Assay, Electron Microscopy

A PH-domain probe indicates that sites of cell wall invaginations correlate with increased PI 4,5 P 2 . (A) Indicated cell types carrying the Cdc10 septin fused to GFP (Cdc10-GFP) and the PLCδ1 PH domain fused to RFP (PH-RFP) were analyzed by fluorescence microscopy. Note the similar localization of these PI 4,5 P 2 -binding proteins. (B) Cell walls were stained with calcofluor white to detect abnormalities. The fraction of cells displaying strongly abnormal cell wall was higher in the sur7∆ and pil1∆ lsp1∆ mutants than in the inp51∆ mutant, which lacks this PI 4,5 P 2 phosphatase. Bottom, representative samples of cells that were scored as wild type, minor defect, or major defect. The contrast was adjusted when scoring samples to make minor defects more obvious. (C) Dilutions of cells were spotted onto rich YPD medium in the absence and presence of 35 μg/ml calcofluor white to perturb the cell wall. Two independent inp51∆ mutants. Strains used were CDC10-GFP PH-RFP (YHXW47-1), CDC10-GFP PH-RFP sur7∆ (YHXW41-1), CDC10-GFP PH-RFP pil1∆ lsp1∆ (YHXW39-1), WT control (DIC185), inp51∆-1 (YXH42-1), inp51∆-2 (YXH42-2), sur7∆ (YJA11), and pil1∆ lsp1∆ (YHXW21-1). Bars, 5 μm.
Figure Legend Snippet: A PH-domain probe indicates that sites of cell wall invaginations correlate with increased PI 4,5 P 2 . (A) Indicated cell types carrying the Cdc10 septin fused to GFP (Cdc10-GFP) and the PLCδ1 PH domain fused to RFP (PH-RFP) were analyzed by fluorescence microscopy. Note the similar localization of these PI 4,5 P 2 -binding proteins. (B) Cell walls were stained with calcofluor white to detect abnormalities. The fraction of cells displaying strongly abnormal cell wall was higher in the sur7∆ and pil1∆ lsp1∆ mutants than in the inp51∆ mutant, which lacks this PI 4,5 P 2 phosphatase. Bottom, representative samples of cells that were scored as wild type, minor defect, or major defect. The contrast was adjusted when scoring samples to make minor defects more obvious. (C) Dilutions of cells were spotted onto rich YPD medium in the absence and presence of 35 μg/ml calcofluor white to perturb the cell wall. Two independent inp51∆ mutants. Strains used were CDC10-GFP PH-RFP (YHXW47-1), CDC10-GFP PH-RFP sur7∆ (YHXW41-1), CDC10-GFP PH-RFP pil1∆ lsp1∆ (YHXW39-1), WT control (DIC185), inp51∆-1 (YXH42-1), inp51∆-2 (YXH42-2), sur7∆ (YJA11), and pil1∆ lsp1∆ (YHXW21-1). Bars, 5 μm.

Techniques Used: Fluorescence, Microscopy, Binding Assay, Staining, Mutagenesis

15) Product Images from "Transcription Factor RFX1 Is Crucial for Maintenance of Genome Integrity in Fusarium graminearum"

Article Title: Transcription Factor RFX1 Is Crucial for Maintenance of Genome Integrity in Fusarium graminearum

Journal: Eukaryotic Cell

doi: 10.1128/EC.00293-13

Nucleus and septum formation in hyphae. Histone H1 was tagged with green fluorescent protein (GFP) or red fluorescent protein (RFP) to visualize nuclei. Cell wall was stained with calcofluor white. Arrowheads indicate septa, and arrows indicate micronuclei. Both deletion of rfx1 and bleomycin treatment inhibited septum formation and produced micronuclei in hyphae. WT, wild type; WT+BLM, wild-type hyphae grown with 20 mU/ml bleomycin; DIC, differential interference contrast; Nuclei Septa, overlays of calcofluor white staining and fluorescent protein images. Scale bar = 20 μm.
Figure Legend Snippet: Nucleus and septum formation in hyphae. Histone H1 was tagged with green fluorescent protein (GFP) or red fluorescent protein (RFP) to visualize nuclei. Cell wall was stained with calcofluor white. Arrowheads indicate septa, and arrows indicate micronuclei. Both deletion of rfx1 and bleomycin treatment inhibited septum formation and produced micronuclei in hyphae. WT, wild type; WT+BLM, wild-type hyphae grown with 20 mU/ml bleomycin; DIC, differential interference contrast; Nuclei Septa, overlays of calcofluor white staining and fluorescent protein images. Scale bar = 20 μm.

Techniques Used: Staining, Produced

16) Product Images from "Helicobacter pylori Biofilm Formation Is Differentially Affected by Common Culture Conditions, and Proteins Play a Central Role in the Biofilm Matrix"

Article Title: Helicobacter pylori Biofilm Formation Is Differentially Affected by Common Culture Conditions, and Proteins Play a Central Role in the Biofilm Matrix

Journal: Applied and Environmental Microbiology

doi: 10.1128/AEM.00391-18

Visualization of eDNA, protein, and polysaccharides in the extracellular matrix of H. pylori biofilms. Staining of the H. pylori biofilm matrix at 24, 48, and 72 h. Representative regions for areas with pronounced biofilm formation are shown. Bar, 100 μm (scale is identical for all panels). Each dye was captured on a single channel, and the images show a compressed z-stack; merged images at a magnification of ×40 are shown on the far right. Biofilms were stained with H. pylori -specific antibodies labeled with Alexa Fluor 488 ( H. pylori ), propidium iodide (DNA), FilmTracer Sypro Ruby biofilm matrix stain (protein), and calcofluor white (polysaccharides).
Figure Legend Snippet: Visualization of eDNA, protein, and polysaccharides in the extracellular matrix of H. pylori biofilms. Staining of the H. pylori biofilm matrix at 24, 48, and 72 h. Representative regions for areas with pronounced biofilm formation are shown. Bar, 100 μm (scale is identical for all panels). Each dye was captured on a single channel, and the images show a compressed z-stack; merged images at a magnification of ×40 are shown on the far right. Biofilms were stained with H. pylori -specific antibodies labeled with Alexa Fluor 488 ( H. pylori ), propidium iodide (DNA), FilmTracer Sypro Ruby biofilm matrix stain (protein), and calcofluor white (polysaccharides).

Techniques Used: Staining, Labeling

17) Product Images from "The Cell Wall Protein Ecm33 of Candida albicans is Involved in Chronological Life Span, Morphogenesis, Cell Wall Regeneration, Stress Tolerance, and Host–Cell Interaction"

Article Title: The Cell Wall Protein Ecm33 of Candida albicans is Involved in Chronological Life Span, Morphogenesis, Cell Wall Regeneration, Stress Tolerance, and Host–Cell Interaction

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2016.00064

Ecm33 is important for nuclear DNA division, septa distribution and actin polarization during C. albicans growth. (A) calcofluor staining was used to visualize the chitin localized in the septum. (B) The nuclear DNA was observed by labeling with DAPI, a highly specific stain for DNA. Arrows indicate defects in the number of nuclei. (C) C. albicans strains were fixed and stained with Alexa phalloidin to visualize actin. Arrows in SC5314 indicate normal actin polarization in buds, compared to RML2U.
Figure Legend Snippet: Ecm33 is important for nuclear DNA division, septa distribution and actin polarization during C. albicans growth. (A) calcofluor staining was used to visualize the chitin localized in the septum. (B) The nuclear DNA was observed by labeling with DAPI, a highly specific stain for DNA. Arrows indicate defects in the number of nuclei. (C) C. albicans strains were fixed and stained with Alexa phalloidin to visualize actin. Arrows in SC5314 indicate normal actin polarization in buds, compared to RML2U.

Techniques Used: Staining, Labeling

18) Product Images from "The exocyst subunit Sec3 is regulated by a protein quality control pathway"

Article Title: The exocyst subunit Sec3 is regulated by a protein quality control pathway

Journal: The Journal of Biological Chemistry

doi: 10.1074/jbc.M117.789867

Hsp70-type molecular chaperones regulate Sec3-913 levels. A , growth on rich medium of the indicated mutant and double mutant strains was compared over a range of temperatures. Note that the temperature-sensitive growth defect of the sec3-913 single mutant is suppressed in the sec3-913sks2 Δ and sec3-913ssa1 Δ strains. B , calcofluor staining of sec3-913 , sec3-913sks2 Δ, and sec3-913ssa1 Δ cells at 30 °C and 34 °C ( left panel ). Scale bar = 5 μm. The percentage of septated cells ( septation index ) and of septated cells with multiple septa were determined. The error bars indicate the standard deviation ( n = 4). *, p
Figure Legend Snippet: Hsp70-type molecular chaperones regulate Sec3-913 levels. A , growth on rich medium of the indicated mutant and double mutant strains was compared over a range of temperatures. Note that the temperature-sensitive growth defect of the sec3-913 single mutant is suppressed in the sec3-913sks2 Δ and sec3-913ssa1 Δ strains. B , calcofluor staining of sec3-913 , sec3-913sks2 Δ, and sec3-913ssa1 Δ cells at 30 °C and 34 °C ( left panel ). Scale bar = 5 μm. The percentage of septated cells ( septation index ) and of septated cells with multiple septa were determined. The error bars indicate the standard deviation ( n = 4). *, p

Techniques Used: Mutagenesis, Staining, Standard Deviation

Deletion of nas6 restores Sec3-913 function in endocytosis and exocytosis. A , calcofluor staining of sec3-913 and sec3-913nas6 Δ cells at 30 °C and 34 °C ( left panel ). The percentages of septated cells ( septation index ) and of septated cells with multiple septa were determined. The error bars indicate the standard deviation ( n = 4). *, p
Figure Legend Snippet: Deletion of nas6 restores Sec3-913 function in endocytosis and exocytosis. A , calcofluor staining of sec3-913 and sec3-913nas6 Δ cells at 30 °C and 34 °C ( left panel ). The percentages of septated cells ( septation index ) and of septated cells with multiple septa were determined. The error bars indicate the standard deviation ( n = 4). *, p

Techniques Used: Staining, Standard Deviation

Sec3-913 is a target of proteasome-associated deubiquitylating enzymes. A , growth on rich medium of the indicated mutant and double mutant strains was compared over a range of temperatures. Note that the temperature-sensitive growth defect of the sec3-913 single mutant is suppressed in sec3-913ubp3 Δ cells and, to a lesser degree, in the sec3-913uch2 Δ strain. B , calcofluor staining of sec3-913 and sec3-913ubp3 Δ cells at 30 °C and 34 °C ( left panel ). Scale bar = 5 μm. The percentages of septated cells ( septation index ) and of septated cells with multiple septa were determined. The error bars indicate the standard deviation ( n = 4). ***, p
Figure Legend Snippet: Sec3-913 is a target of proteasome-associated deubiquitylating enzymes. A , growth on rich medium of the indicated mutant and double mutant strains was compared over a range of temperatures. Note that the temperature-sensitive growth defect of the sec3-913 single mutant is suppressed in sec3-913ubp3 Δ cells and, to a lesser degree, in the sec3-913uch2 Δ strain. B , calcofluor staining of sec3-913 and sec3-913ubp3 Δ cells at 30 °C and 34 °C ( left panel ). Scale bar = 5 μm. The percentages of septated cells ( septation index ) and of septated cells with multiple septa were determined. The error bars indicate the standard deviation ( n = 4). ***, p

Techniques Used: Mutagenesis, Staining, Standard Deviation

19) Product Images from "The first isolate of Candida auris in China: clinical and biological aspects"

Article Title: The first isolate of Candida auris in China: clinical and biological aspects

Journal: Emerging Microbes & Infections

doi: 10.1038/s41426-018-0095-0

Morphologies of C . auris cells grown on YPD ( a ) and YPD plus 10% NaCl ( b ) media. Cells (1 × 10 5 ) were spotted onto different medias and cultured at 37 °C and 40 °C for five days. Cells were collected and stained with DAPI or Calcofluor white. Scale bar, 10 μm. DIC, differential interference contrast
Figure Legend Snippet: Morphologies of C . auris cells grown on YPD ( a ) and YPD plus 10% NaCl ( b ) media. Cells (1 × 10 5 ) were spotted onto different medias and cultured at 37 °C and 40 °C for five days. Cells were collected and stained with DAPI or Calcofluor white. Scale bar, 10 μm. DIC, differential interference contrast

Techniques Used: Cell Culture, Staining

20) Product Images from "The polyketide synthase PKS15 has a crucial role in cell wall formation in Beauveria bassiana"

Article Title: The polyketide synthase PKS15 has a crucial role in cell wall formation in Beauveria bassiana

Journal: Scientific Reports

doi: 10.1038/s41598-020-69417-w

Size and shape of fluorescently-stained conidia in the wild type (WT), Δ pks15 and the complemented isolate G6. ( a ) Calcofluor- and FITC-tagged concanavalin A staining (upper and lower panels). Bars, 5 μm. ( b ) Distribution of sizes in the conidial populations of the wild type, Δ pks15 and G6 from a single representative experiment. ( c ) Frequency of conidial sizes for each of the three strains. Size data were from three independent experiments. ( d ) Distribution of shapes in the conidial populations of wild type, Δ pks15 and G6 from a single representative experiment. Shape factor (circularity) was determined using the NIS-Elements D software. A shape factor of 1.0 indicates a circle, whereas a shape factor less than 1.0 indicates an ellipse. ( e ) Frequency of conidial shapes for each of the three strains. Data shown are mean ± s.e.m. Asterisks indicate statistical significance between the wild type or the complemented isolate G6 and Δ pks15 (Student’s t -test: * p
Figure Legend Snippet: Size and shape of fluorescently-stained conidia in the wild type (WT), Δ pks15 and the complemented isolate G6. ( a ) Calcofluor- and FITC-tagged concanavalin A staining (upper and lower panels). Bars, 5 μm. ( b ) Distribution of sizes in the conidial populations of the wild type, Δ pks15 and G6 from a single representative experiment. ( c ) Frequency of conidial sizes for each of the three strains. Size data were from three independent experiments. ( d ) Distribution of shapes in the conidial populations of wild type, Δ pks15 and G6 from a single representative experiment. Shape factor (circularity) was determined using the NIS-Elements D software. A shape factor of 1.0 indicates a circle, whereas a shape factor less than 1.0 indicates an ellipse. ( e ) Frequency of conidial shapes for each of the three strains. Data shown are mean ± s.e.m. Asterisks indicate statistical significance between the wild type or the complemented isolate G6 and Δ pks15 (Student’s t -test: * p

Techniques Used: Staining, Software

21) Product Images from "A unicellular relative of animals generates a layer of polarized cells by actomyosin-dependent cellularization"

Article Title: A unicellular relative of animals generates a layer of polarized cells by actomyosin-dependent cellularization

Journal: eLife

doi: 10.7554/eLife.49801

Actin cytoskeleton, plasma membrane and cell-wall dynamics during cellularization of S. arctica. ( A ) Rate of membrane invagination during cellularization reveal the presence of an early slow phase followed by a fast one. Measurements were obtained from 15 FM4-64 stained independent coenocytes. For the measurements of each cell, a mean was obtained from three single invaginations. Traces from single cells are represented in dashed lines whereas the mean of all 15 cells is shown in black with its corresponding standard deviation. ( B ) Epifluorescence microscopy images of the cellularization process using phalloidin and DAPI. Arrows show the different actin structures during the different stages of cellularization. Bars, 50 µm for whole image and 10 µm for the zoom. ( C ) fraction of cells exhibiting different actin and cellular structures throughout cellularization. The temporal order of actin structures show that the stages occur sequentially. (N°cells = 300). ( D ) Cell wall formation is observed only post-flip. Cells were fixed and co-stained with the membrane dye FM4-64FX and the cell-wall dye calcofluor white and imaged using an epifluorescence microscope. Bar, 10 µm.
Figure Legend Snippet: Actin cytoskeleton, plasma membrane and cell-wall dynamics during cellularization of S. arctica. ( A ) Rate of membrane invagination during cellularization reveal the presence of an early slow phase followed by a fast one. Measurements were obtained from 15 FM4-64 stained independent coenocytes. For the measurements of each cell, a mean was obtained from three single invaginations. Traces from single cells are represented in dashed lines whereas the mean of all 15 cells is shown in black with its corresponding standard deviation. ( B ) Epifluorescence microscopy images of the cellularization process using phalloidin and DAPI. Arrows show the different actin structures during the different stages of cellularization. Bars, 50 µm for whole image and 10 µm for the zoom. ( C ) fraction of cells exhibiting different actin and cellular structures throughout cellularization. The temporal order of actin structures show that the stages occur sequentially. (N°cells = 300). ( D ) Cell wall formation is observed only post-flip. Cells were fixed and co-stained with the membrane dye FM4-64FX and the cell-wall dye calcofluor white and imaged using an epifluorescence microscope. Bar, 10 µm.

Techniques Used: Staining, Standard Deviation, Epifluorescence Microscopy, Microscopy

22) Product Images from "The first isolate of Candida auris in China: clinical and biological aspects"

Article Title: The first isolate of Candida auris in China: clinical and biological aspects

Journal: Emerging Microbes & Infections

doi: 10.1038/s41426-018-0095-0

Morphologies of C . auris cells grown on YPD ( a ) and YPD plus 10% NaCl ( b ) media. Cells (1 × 10 5 ) were spotted onto different medias and cultured at 37 °C and 40 °C for five days. Cells were collected and stained with DAPI or Calcofluor white. Scale bar, 10 μm. DIC, differential interference contrast
Figure Legend Snippet: Morphologies of C . auris cells grown on YPD ( a ) and YPD plus 10% NaCl ( b ) media. Cells (1 × 10 5 ) were spotted onto different medias and cultured at 37 °C and 40 °C for five days. Cells were collected and stained with DAPI or Calcofluor white. Scale bar, 10 μm. DIC, differential interference contrast

Techniques Used: Cell Culture, Staining

23) Product Images from "Signaling Domains of Mucin Msb2 in Candida albicans"

Article Title: Signaling Domains of Mucin Msb2 in Candida albicans

Journal: Eukaryotic Cell

doi: 10.1128/EC.00264-14

Msb2-Sho1 interactions induce hypha formation via Cek1 and Efg1. (A) Hyperfilamentous growth of strains producing Msb2 protein variants ΔD11 and ΔD12. Transformants (encoded variants) included strains ESCa25 (ΔD10), MSCa37 (ΔD11), MSCa45 (ΔD12), MSCa46 (ΔD13), and MSCa48 (ΔD9). (B) Hyphal induction induced by the ΔD11 Msb2 variant in different genetic backgrounds. Resulting transformants included MSCa37 ( msb2 [ ACT1p-MSB2 ΔN 514–1087 ]), MSCa38 (wt [ ACT1p-MSB2 ΔN 514–1087 ]), MSCa39 ( cek1 [ ACT1p-MSB2 ΔN 514–1087 ]), MSCa40 ( czf1 [ ACT1p-MSB2 ΔN 514–1087 ]), MSCa41 ( efg1 [ ACT1p-MSB2 ΔN 514–1087 ]), MSCa42 ( cph1 [ ACT1p-MSB2 ΔN 514–1087 ]), MSCa43 ( tpk1 [ ACT1p-MSB2 ΔN 514–1087 ]), MSCa44 ( tpk2 [ ACT1p-MSB2 ΔN 514–1087 ]), and MSCa47 ( msb2 sho1 [ ACT1p-MSB2 ΔN 514–1087 ]). All cells were grown at 30°C in YPD before staining with calcofluor white and microscopic inspection.
Figure Legend Snippet: Msb2-Sho1 interactions induce hypha formation via Cek1 and Efg1. (A) Hyperfilamentous growth of strains producing Msb2 protein variants ΔD11 and ΔD12. Transformants (encoded variants) included strains ESCa25 (ΔD10), MSCa37 (ΔD11), MSCa45 (ΔD12), MSCa46 (ΔD13), and MSCa48 (ΔD9). (B) Hyphal induction induced by the ΔD11 Msb2 variant in different genetic backgrounds. Resulting transformants included MSCa37 ( msb2 [ ACT1p-MSB2 ΔN 514–1087 ]), MSCa38 (wt [ ACT1p-MSB2 ΔN 514–1087 ]), MSCa39 ( cek1 [ ACT1p-MSB2 ΔN 514–1087 ]), MSCa40 ( czf1 [ ACT1p-MSB2 ΔN 514–1087 ]), MSCa41 ( efg1 [ ACT1p-MSB2 ΔN 514–1087 ]), MSCa42 ( cph1 [ ACT1p-MSB2 ΔN 514–1087 ]), MSCa43 ( tpk1 [ ACT1p-MSB2 ΔN 514–1087 ]), MSCa44 ( tpk2 [ ACT1p-MSB2 ΔN 514–1087 ]), and MSCa47 ( msb2 sho1 [ ACT1p-MSB2 ΔN 514–1087 ]). All cells were grown at 30°C in YPD before staining with calcofluor white and microscopic inspection.

Techniques Used: Variant Assay, Staining

24) Product Images from "Determination of ACC-induced cell-programmed death in roots of Vicia faba ssp. minor seedlings by acridine orange and ethidium bromide staining"

Article Title: Determination of ACC-induced cell-programmed death in roots of Vicia faba ssp. minor seedlings by acridine orange and ethidium bromide staining

Journal: Protoplasma

doi: 10.1007/s00709-012-0383-9

Cross section of the cortex of V. faba ssp. minor seedling roots with ACC-induced aerenchyma spaces (indicated by arrows ), at the early stages of its formation, visualized under a white light ( a ), fluorescence, after cellulose with calcofluor white staining, ( b ) microscope, confirmed by plot profile ( c ) and surface plot ( d ). Scale bars are 100 μm
Figure Legend Snippet: Cross section of the cortex of V. faba ssp. minor seedling roots with ACC-induced aerenchyma spaces (indicated by arrows ), at the early stages of its formation, visualized under a white light ( a ), fluorescence, after cellulose with calcofluor white staining, ( b ) microscope, confirmed by plot profile ( c ) and surface plot ( d ). Scale bars are 100 μm

Techniques Used: Fluorescence, Staining, Microscopy

25) Product Images from "Transcript Profiles of Candida albicans Cortical Actin Patch Mutants Reflect Their Cellular Defects: Contribution of the Hog1p and Mkc1p Signaling Pathways †"

Article Title: Transcript Profiles of Candida albicans Cortical Actin Patch Mutants Reflect Their Cellular Defects: Contribution of the Hog1p and Mkc1p Signaling Pathways †

Journal:

doi: 10.1128/EC.00385-05

Aberrant chitin depositions in the cell walls of myo5 Δ and sla2 Δ mutant cells. Wild-type and mutant cells were fixed, stained with calcofluor white, and visualized by epifluorescence microscopy. Bar = 5 μm.
Figure Legend Snippet: Aberrant chitin depositions in the cell walls of myo5 Δ and sla2 Δ mutant cells. Wild-type and mutant cells were fixed, stained with calcofluor white, and visualized by epifluorescence microscopy. Bar = 5 μm.

Techniques Used: Mutagenesis, Staining, Epifluorescence Microscopy

26) Product Images from "Heterogeneous distribution of Candida albicans cell-surface antigens demonstrated with an Als1-specific monoclonal antibody"

Article Title: Heterogeneous distribution of Candida albicans cell-surface antigens demonstrated with an Als1-specific monoclonal antibody

Journal: Microbiology

doi: 10.1099/mic.0.043851-0

C. albicans yeast and germ tubes overexpressing ALS1 have a uniform cell-surface coat of Als1, even within bud scars. Strain 2243, which overexpressed ALS1 under the control of the constitutive, high-activity TPI1 promoter, was grown in YPD for 16 h at 30 °C and transferred to fresh culture medium. (a) Yeast cells from 8 h growth at 30 °C in fresh YPD medium were fixed in paraformaldehyde and immunolabelled with anti-Als1 and a FITC-conjugated secondary antibody. In contrast to the labelling pattern of the control C. albicans strain CAI12 (Fig. 1 ), yeast forms of the ALS1 overexpression strain were labelled strongly at the poles of the cell, suggesting Als1 localization within bud scars. This conclusion was strengthened by observation of overlapping fluorescent signals from anti-Als1 immunolabelling and Calcofluor White staining (b). From left, a Calcofluor-stained cell false coloured with cyan, FITC anti-Als1 labelling false coloured yellow, the merged image, and a bright-field image. (c) YPD-grown 16 h yeast cells of strain 2243 were inoculated into RPMI medium for 1 h to produce germ tubes. Strong anti-Als1 immunolabelling was observed over the surface of the mother yeast and the entire germ tube. All images were produced using a Zeiss LSM 710 microscope.
Figure Legend Snippet: C. albicans yeast and germ tubes overexpressing ALS1 have a uniform cell-surface coat of Als1, even within bud scars. Strain 2243, which overexpressed ALS1 under the control of the constitutive, high-activity TPI1 promoter, was grown in YPD for 16 h at 30 °C and transferred to fresh culture medium. (a) Yeast cells from 8 h growth at 30 °C in fresh YPD medium were fixed in paraformaldehyde and immunolabelled with anti-Als1 and a FITC-conjugated secondary antibody. In contrast to the labelling pattern of the control C. albicans strain CAI12 (Fig. 1 ), yeast forms of the ALS1 overexpression strain were labelled strongly at the poles of the cell, suggesting Als1 localization within bud scars. This conclusion was strengthened by observation of overlapping fluorescent signals from anti-Als1 immunolabelling and Calcofluor White staining (b). From left, a Calcofluor-stained cell false coloured with cyan, FITC anti-Als1 labelling false coloured yellow, the merged image, and a bright-field image. (c) YPD-grown 16 h yeast cells of strain 2243 were inoculated into RPMI medium for 1 h to produce germ tubes. Strong anti-Als1 immunolabelling was observed over the surface of the mother yeast and the entire germ tube. All images were produced using a Zeiss LSM 710 microscope.

Techniques Used: Activity Assay, Over Expression, Staining, Produced, Microscopy

Anti-Als1 immunolabelling and Calcofluor White staining of C. albicans yeast cells to demonstrate the absence of Als1 localization to the bud scar. C. albicans strain CAI12 from a saturated culture was grown for 1 h in fresh YPD medium at 30 °C and fixed in paraformaldehyde. Fixed cells were immunolabelled with anti-Als1 and a FITC-conjugated secondary antibody. Cells were then stained with Calcofluor White, which binds to chitin, accentuating the fungal cell wall and bud scars. Cells were visualized with a Zeiss LSM 710 microscope. Rows (a–d) show images of Calcofluor-stained cells (left column, false coloured with cyan), FITC anti-Als1 labelling (second column from left, false coloured with yellow), a merged image (second column from right) and a bright-field image (right column). FITC labelling demonstrated Als1 on the cell surface with the exception of the bud scars, which were clearly stained with Calcofluor. Arrows within each bright-field image denote the plane of focus: an arrow with a tail shows cells with edges in focus, while arrowheads show cells where the cell surface was in focus so that the bud scar structure could be viewed more readily.
Figure Legend Snippet: Anti-Als1 immunolabelling and Calcofluor White staining of C. albicans yeast cells to demonstrate the absence of Als1 localization to the bud scar. C. albicans strain CAI12 from a saturated culture was grown for 1 h in fresh YPD medium at 30 °C and fixed in paraformaldehyde. Fixed cells were immunolabelled with anti-Als1 and a FITC-conjugated secondary antibody. Cells were then stained with Calcofluor White, which binds to chitin, accentuating the fungal cell wall and bud scars. Cells were visualized with a Zeiss LSM 710 microscope. Rows (a–d) show images of Calcofluor-stained cells (left column, false coloured with cyan), FITC anti-Als1 labelling (second column from left, false coloured with yellow), a merged image (second column from right) and a bright-field image (right column). FITC labelling demonstrated Als1 on the cell surface with the exception of the bud scars, which were clearly stained with Calcofluor. Arrows within each bright-field image denote the plane of focus: an arrow with a tail shows cells with edges in focus, while arrowheads show cells where the cell surface was in focus so that the bud scar structure could be viewed more readily.

Techniques Used: Staining, Microscopy

27) Product Images from "Structure-function analysis of the maize bulliform cell cuticle and its role in dehydration and leaf rolling"

Article Title: Structure-function analysis of the maize bulliform cell cuticle and its role in dehydration and leaf rolling

Journal: bioRxiv

doi: 10.1101/2020.02.06.937011

Three different epidermal mutants have increased bulliform cell coverage. A-D) Bright field images of hand-sectioned adult leaves from the indicated genotypes depict previously reported aberrations in epidermal cell types. Arrow = wild-type-like bulliform strip (2 cells wide), arrowhead = areas with abnormal bulliform-like cells. Scale bar = 200 µm. E) Quantification of bulliform cell number in three bulliform mutants as percentage of all epidermal cells, calculated using (F-I) Fluorol Yellow stained tissue cross section images. FY = Fluorol Yellow (lipid stain; green), CW = Calcofluor White (cell wall counter stain; blue). White arrow = FY staining of wild-type-like bulliform strip cuticle (3 cells wide), white arrowhead in G-I) = areas with abnormal BC-like cells displaying increased FY staining of the cuticle. Scale bar = 50 µm. Values in E) are given as means ± SD (n = 300-380 epidermal cells counted, 4 biological replicates per genotype). Statistical analysis used two-tailed unpaired Student’s t -test, with *P
Figure Legend Snippet: Three different epidermal mutants have increased bulliform cell coverage. A-D) Bright field images of hand-sectioned adult leaves from the indicated genotypes depict previously reported aberrations in epidermal cell types. Arrow = wild-type-like bulliform strip (2 cells wide), arrowhead = areas with abnormal bulliform-like cells. Scale bar = 200 µm. E) Quantification of bulliform cell number in three bulliform mutants as percentage of all epidermal cells, calculated using (F-I) Fluorol Yellow stained tissue cross section images. FY = Fluorol Yellow (lipid stain; green), CW = Calcofluor White (cell wall counter stain; blue). White arrow = FY staining of wild-type-like bulliform strip cuticle (3 cells wide), white arrowhead in G-I) = areas with abnormal BC-like cells displaying increased FY staining of the cuticle. Scale bar = 50 µm. Values in E) are given as means ± SD (n = 300-380 epidermal cells counted, 4 biological replicates per genotype). Statistical analysis used two-tailed unpaired Student’s t -test, with *P

Techniques Used: Stripping Membranes, Staining, Two Tailed Test

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  • 99
    Millipore calcofluor white m2r cfw
    Three-dimensional projections of individual components in biofilms formed by four methicillin- resistant Staphylococcus aureus (MRSA) strains on polystyrene after 24 h of incubation. The images were obtained from confocal stack images by the IMARIS 9.1 software, virtual projections being included on the right. 1 propidium iodide; 2 fluorescein isothiocyanate isomer I; 3 DiIC18(5) oil, 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindodicarbocyanine perchlorate; 4 concanavalin A, tetramethylrhodamine conjugate; 5 <t>calcofluor</t> white <t>M2R.</t>
    Calcofluor White M2r Cfw, supplied by Millipore, used in various techniques. Bioz Stars score: 99/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    98
    Millipore calcofluor white
    PKC1 is vital for protection against cell wall stress. Cultures of KN99 a and the pkc1Δ and complemented pck1Δ :: PKC1 strains were grown overnight in YPD containing 1 M sorbitol at 30°C and then diluted to an OD 650 of 1.0. Five-microliter portions of 10-fold serial dilutions were plated onto YPD plates containing 1 M sorbitol plus the indicated inhibitor and grown for 3 days at 30°C. CFW, <t>calcofluor</t> white.
    Calcofluor White, supplied by Millipore, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Three-dimensional projections of individual components in biofilms formed by four methicillin- resistant Staphylococcus aureus (MRSA) strains on polystyrene after 24 h of incubation. The images were obtained from confocal stack images by the IMARIS 9.1 software, virtual projections being included on the right. 1 propidium iodide; 2 fluorescein isothiocyanate isomer I; 3 DiIC18(5) oil, 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindodicarbocyanine perchlorate; 4 concanavalin A, tetramethylrhodamine conjugate; 5 calcofluor white M2R.

    Journal: Frontiers in Microbiology

    Article Title: Characterization of Biofilms Formed by Foodborne Methicillin-Resistant Staphylococcus aureus

    doi: 10.3389/fmicb.2018.03004

    Figure Lengend Snippet: Three-dimensional projections of individual components in biofilms formed by four methicillin- resistant Staphylococcus aureus (MRSA) strains on polystyrene after 24 h of incubation. The images were obtained from confocal stack images by the IMARIS 9.1 software, virtual projections being included on the right. 1 propidium iodide; 2 fluorescein isothiocyanate isomer I; 3 DiIC18(5) oil, 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindodicarbocyanine perchlorate; 4 concanavalin A, tetramethylrhodamine conjugate; 5 calcofluor white M2R.

    Article Snippet: SYTO 9 and propidium iodide (PI) from the LIVE/DEAD® BacLightTM Bacterial Viability Kit, DiIC18(5) oil, 1,1′-dioctadecyl-3,3,3′,3′-tetramethylindodicarbocyanine perchlorate (DiD’oil) and concanavalin A, tetramethylrhodamine conjugate (ConA-TMR) were purchased from Invitrogen (Carlsbad, CA, United States), while fluorescein isothiocyanate isomer I (FITC) and calcofluor white M2R (CFW) were purchased from Sigma (St. Louis, MO, United States).

    Techniques: Incubation, Software

    Deletion of Fkh2 in B. bassiana affects cell cycle and septation pattern of multicullular hyphae and transcriptional profiles of related genes. ( A ) Bright and fluorescent images of hyphae stained with the cell-wall-specific dye calcofluor white (upper row) and the nucleic-acid-specific dye DAPI (lower row). Scale bars: 10 μm. ( B , C ) Relative transcript levels (RTL) of cell cycle and septation related genes in the 3-day-old SDB cultures of Fkh2 mutants versus WT, respectively. The asterisked bar in each three-bar group differ significantly from two others unmarked (Tukey’s HSD, P

    Journal: Scientific Reports

    Article Title: Transcriptional control of fungal cell cycle and cellular events by Fkh2, a forkhead transcription factor in an insect pathogen

    doi: 10.1038/srep10108

    Figure Lengend Snippet: Deletion of Fkh2 in B. bassiana affects cell cycle and septation pattern of multicullular hyphae and transcriptional profiles of related genes. ( A ) Bright and fluorescent images of hyphae stained with the cell-wall-specific dye calcofluor white (upper row) and the nucleic-acid-specific dye DAPI (lower row). Scale bars: 10 μm. ( B , C ) Relative transcript levels (RTL) of cell cycle and septation related genes in the 3-day-old SDB cultures of Fkh2 mutants versus WT, respectively. The asterisked bar in each three-bar group differ significantly from two others unmarked (Tukey’s HSD, P

    Article Snippet: Hyphae collected from the liquid culture were stained with calcofluor white and DAPI (Sigma) for 15 min, followed by visualization in the same bright/fluorescent field under a confocal microscope.

    Techniques: Staining

    Effect of PHBs on the monokaryotic S1–10 strain of Lentinula edodes . Expression of the downstream genes in the mating pathway was monitored time-dependently. PHB1 or PHB9 (20 μg/mL) was added to actively growing mycelia of L. edodes S1–10 strain at 25 °C. Inset pictures show the microscopic image of the mushroom mycelia after the treatment of PHB1 or PHB9. The mycelia were stained with calcofluor. The circles indicate clamp connections. Error bars indicate standard deviations of means. The statistical significance of the mean difference is indicated on the bar with asterisks (** for p ≤ 0.01, and **** for p ≤ 0.0001).

    Journal: Genes

    Article Title: Investigation of Mating Pheromone–Pheromone Receptor Specificity in Lentinula edodes

    doi: 10.3390/genes11050506

    Figure Lengend Snippet: Effect of PHBs on the monokaryotic S1–10 strain of Lentinula edodes . Expression of the downstream genes in the mating pathway was monitored time-dependently. PHB1 or PHB9 (20 μg/mL) was added to actively growing mycelia of L. edodes S1–10 strain at 25 °C. Inset pictures show the microscopic image of the mushroom mycelia after the treatment of PHB1 or PHB9. The mycelia were stained with calcofluor. The circles indicate clamp connections. Error bars indicate standard deviations of means. The statistical significance of the mean difference is indicated on the bar with asterisks (** for p ≤ 0.01, and **** for p ≤ 0.0001).

    Article Snippet: For the microscopic analysis, the PHB-treated mycelia were stained with equivalent mixture of 10% potassium hydroxide and calcofluor white (Sigma-Aldrich), which binds to the chitin component of the fungal cell wall.

    Techniques: Expressing, Staining

    PKC1 is vital for protection against cell wall stress. Cultures of KN99 a and the pkc1Δ and complemented pck1Δ :: PKC1 strains were grown overnight in YPD containing 1 M sorbitol at 30°C and then diluted to an OD 650 of 1.0. Five-microliter portions of 10-fold serial dilutions were plated onto YPD plates containing 1 M sorbitol plus the indicated inhibitor and grown for 3 days at 30°C. CFW, calcofluor white.

    Journal: Eukaryotic Cell

    Article Title: PKC1 Is Essential for Protection against both Oxidative and Nitrosative Stresses, Cell Integrity, and Normal Manifestation of Virulence Factors in the Pathogenic Fungus Cryptococcus neoformans ▿ ▿ †

    doi: 10.1128/EC.00146-08

    Figure Lengend Snippet: PKC1 is vital for protection against cell wall stress. Cultures of KN99 a and the pkc1Δ and complemented pck1Δ :: PKC1 strains were grown overnight in YPD containing 1 M sorbitol at 30°C and then diluted to an OD 650 of 1.0. Five-microliter portions of 10-fold serial dilutions were plated onto YPD plates containing 1 M sorbitol plus the indicated inhibitor and grown for 3 days at 30°C. CFW, calcofluor white.

    Article Snippet: Tenfold serial dilutions were made in PBS plus 1 M sorbitol and 5 μl of these was plated on YPD plates containing 1 M sorbitol without and with 0.5, 1.0, and 1.5 mg/ml calcofluor white (Fluorescent Brightner 28; Sigma, F-3543); 0.2, 0.5, and 1.0 mg/ml caffeine (Sigma, C-0750); 0.01, 0.03, and 0.06% SDS (AnalaR Biochemical, BDH Chemicals Ltd., Poole, England); 0.5% Congo red (Sigma, C-6767); and 1.5 M NaCl.

    Techniques:

    FLAG-tagged Mpk1 allows for the detection of total Mpk1, the terminal kinase in the PKC1 pathway, independent of phosphorylation. (A) Western blot analysis was done using the anti-Mpk1 phospho-antibody to probe a blot containing total protein from wild-type or mpk1Δ lysates, which were made from cells that were either uninduced or induced with calcofluor white (CFW). The amount of the 50-kDa band increased in the wild-type strain in response to CFW, and the absence of the band in the mpk1Δ lysate indicated that the band in the wild-type lysate was Mpk1. (B) The gene construct encoding the Mpk1 protein fused to the Flag epitope at the carboxy terminus replaced the wild-type MPK1 locus in KN99α. The panel shows Western blot analysis using an antibody against the Flag tag and lysates from the Mpk1-Flag tagged strain and the wild-type strain either induced or uninduced with CFW. Bands of the appropriate size were detected in the Mpk1-Flag strain but not the wild-type strain, suggesting that the protein detected was the Mpk1-Flag fusion. (C) The same blot from panel B probed with an antibody specific for the phosphorylated form of Mpk1. Phosphorylation of both the fusion protein and the wild-type protein was induced by CFW.

    Journal: Eukaryotic Cell

    Article Title: PKC1 Is Essential for Protection against both Oxidative and Nitrosative Stresses, Cell Integrity, and Normal Manifestation of Virulence Factors in the Pathogenic Fungus Cryptococcus neoformans ▿ ▿ †

    doi: 10.1128/EC.00146-08

    Figure Lengend Snippet: FLAG-tagged Mpk1 allows for the detection of total Mpk1, the terminal kinase in the PKC1 pathway, independent of phosphorylation. (A) Western blot analysis was done using the anti-Mpk1 phospho-antibody to probe a blot containing total protein from wild-type or mpk1Δ lysates, which were made from cells that were either uninduced or induced with calcofluor white (CFW). The amount of the 50-kDa band increased in the wild-type strain in response to CFW, and the absence of the band in the mpk1Δ lysate indicated that the band in the wild-type lysate was Mpk1. (B) The gene construct encoding the Mpk1 protein fused to the Flag epitope at the carboxy terminus replaced the wild-type MPK1 locus in KN99α. The panel shows Western blot analysis using an antibody against the Flag tag and lysates from the Mpk1-Flag tagged strain and the wild-type strain either induced or uninduced with CFW. Bands of the appropriate size were detected in the Mpk1-Flag strain but not the wild-type strain, suggesting that the protein detected was the Mpk1-Flag fusion. (C) The same blot from panel B probed with an antibody specific for the phosphorylated form of Mpk1. Phosphorylation of both the fusion protein and the wild-type protein was induced by CFW.

    Article Snippet: Tenfold serial dilutions were made in PBS plus 1 M sorbitol and 5 μl of these was plated on YPD plates containing 1 M sorbitol without and with 0.5, 1.0, and 1.5 mg/ml calcofluor white (Fluorescent Brightner 28; Sigma, F-3543); 0.2, 0.5, and 1.0 mg/ml caffeine (Sigma, C-0750); 0.01, 0.03, and 0.06% SDS (AnalaR Biochemical, BDH Chemicals Ltd., Poole, England); 0.5% Congo red (Sigma, C-6767); and 1.5 M NaCl.

    Techniques: Western Blot, Construct, FLAG-tag