Journal: eLife

Article Title: The Mycobacterium ulcerans toxin mycolactone causes destructive Sec61-dependent loss of the endothelial glycocalyx and vessel basement membrane to drive skin necrosis

doi: 10.7554/eLife.86931

Figure Lengend Snippet:

Article Snippet: Antibody , Anti-integrin β4/CD104 (rat monoclonal 439-9B) , eBioscience , 14-1049-82 RRID: AB_1210460 , FACS (1:100).

Techniques: Plasmid Preparation, Control, Sequencing, Negative Control, Recombinant, Software, Transfection, Histopathology, Western Blot, Staining, Permeability

Journal: bioRxiv

Article Title: Molecular and Mechanical Signatures Contributing to Epidermal Differentiation and Barrier Formation

doi: 10.1101/2024.07.23.604736

Figure Lengend Snippet: (A) Diagram depicting the stratification process and different cell populations (basal and K10+ suprabasal layers) existing in the epidermis from E14.5 to E16.5. (B) Images of EdU staining (white) at E14.5, E15.5 and E16.5. Immunofluorescence of suprabasal cells labeled with K10 in red and the basement membrane, with β4-integrin in green. Scale bars: 20 μm. (C) Quantification of the percentage of EdU+ cells in the basal and the first two layers of suprabasal cells at E14.5, E15.5 and E16.5. n= 3 embryos/time point. Data are presented as mean ± standard deviation (SD). ns: not significant, *: p < 0.5, **: p < 0.01, ***: p < 0.001, ordinary two-way ANOVA, Tukey’s multiple comparisons test. (D) Schematic showing basal and suprabasal cell populations collected for RNA sequencing: K14-RFP;K10-rtTA;TRE-H2B-GFP pregnant dams were all fed with doxycycline from E12.5, and then sacrificed at either E14.5 or E16.5. The E14.5 intermediate cells (K10-GFP + ;K14-RFP - ), E16.5 spinous cells (K10-GFP + ;K14-RFP - ), and basal cells at E14.5 and E16.5 (K10-GFP - ;K14-RFP + ) cells were collected separately and sent for RNA sequencing. Granular cells were excluded from E16.5 samples. (E) PCA score plot of the first two principal components (PC1: 33.5% variance and PC2: 21.1% variance) for gene expression levels from samples of cell populations indicated in (D) (n=3 embryos/cell population).

Article Snippet: Primary antibodies used in this study: rat anti-HA (Sigma-Aldrich, 11867423001), chicken anti-keratin 5/14 (generated in the Lechler lab), rabbit anti-keratin 10 (Covance, 905401), guinea pig anti-keratin 10 (Progen, GP-K10), rat anti-β4 integrin (BD Biosciences, 553745), rabbit anti-MyosinIIA (Biolegend, PRB-440P), rat anti-α18 (gift from Akira Nagafuchi, Kumamoto University), rabbit anti-YAP/TAZ (Cell Signaling Technology, 8418S), rabbit anti-Tgm1(Proteintech,12912-3-AP), rabbit anti-MafB (Novus, NBP1-81342), rabbit anti-St8sia6 (Sigma, HPA011635), rabbit anti-Abca12 (gift from Dr. Wong, University of Michigan).

Techniques: Staining, Immunofluorescence, Labeling, Membrane, Standard Deviation, RNA Sequencing Assay, Expressing

Journal: bioRxiv

Article Title: Molecular and Mechanical Signatures Contributing to Epidermal Differentiation and Barrier Formation

doi: 10.1101/2024.07.23.604736

Figure Lengend Snippet: (A and B) Immunofluorescence staining of Loricrin(A) or St8sia6 (B) in green labeling granular cells at E16.5 but not in intermediate cells at E14.5. Basement membrane is marked with β4-integrin in red. Scale bars: 20 μm.

Article Snippet: Primary antibodies used in this study: rat anti-HA (Sigma-Aldrich, 11867423001), chicken anti-keratin 5/14 (generated in the Lechler lab), rabbit anti-keratin 10 (Covance, 905401), guinea pig anti-keratin 10 (Progen, GP-K10), rat anti-β4 integrin (BD Biosciences, 553745), rabbit anti-MyosinIIA (Biolegend, PRB-440P), rat anti-α18 (gift from Akira Nagafuchi, Kumamoto University), rabbit anti-YAP/TAZ (Cell Signaling Technology, 8418S), rabbit anti-Tgm1(Proteintech,12912-3-AP), rabbit anti-MafB (Novus, NBP1-81342), rabbit anti-St8sia6 (Sigma, HPA011635), rabbit anti-Abca12 (gift from Dr. Wong, University of Michigan).

Techniques: Immunofluorescence, Staining, Labeling, Membrane

Journal: Development (Cambridge, England)

Article Title: A transit-amplifying progenitor with biphasic behavior contributes to epidermal renewal

doi: 10.1242/dev.202389

Figure Lengend Snippet: Aspm is expressed in a distinct subset of highly proliferative basal cells. (A) scRNA-seq feature plots of Sca1 + /α6-integrin + basal layer (BL) cells sorted from mouse tail skin at PD52. *Data extracted from scRNA-seq database. Shades of blue show expression levels. Dlx1 and Aspm mark the BL subsets employed in this study, K14 marks the BL, K10 and Involucrin mark differentiating cells, and Ki67 marks proliferative cells . (B) The scRNA-seq BL data in A were analyzed to determine the fractions of Dlx1 + cells that co-expressed the indicated genes. (C) As in B, except that the Aspm + cells were analyzed. (D) Cell cycle phase PCA analysis of BL cells. (E) Fractions of distinct BL populations in the cell cycle phases. (F,G) Differentially expressed genes (DEGs) that were upregulated (UP) in the indicated cells as analyzed using GO Ontology (2023) identifiers. DEGs in G were extracted by comparing each population with all BL Ki67 + cells. Enrichr combined scores (indicated by color) and gene expression ratio (indicated by ellipse size) are shown. The x -axis in F shows the negative log 10 of the P -value adjusted for multiple testing (Benjamini-Hochberg correction).

Article Snippet: Rabbit anti-K14 (1:100, BioLegend, 905301) or mouse anti-K10 (1:100, BioLegend, 904301) and rat anti-β4-integrin (1:200, BD Biosciences) or rabbit anti-Ki67 (1:100, Abcam, ab15580) were used.

Techniques: Expressing

Journal: Development (Cambridge, England)

Article Title: A transit-amplifying progenitor with biphasic behavior contributes to epidermal renewal

doi: 10.1242/dev.202389

Figure Lengend Snippet: Clonal genetic lineage tracing for Dlx1-CreER and Aspm-CreER epidermal progenitors. (A) Schematic of low dose tamoxifen (TM) induction in mice injected once at PD35 and sacrificed at the chase times indicated. (B) xyz orthogonal projections through optical z -stacks from whole-mount skin collected from the chased mice stained with β4-integrin to mark the basal layer show the basal and suprabasal tdTomato + clones and cells. (C,D) Beeswarm plots of tdTomato + clone and cell counts in images like those shown in B from comparable tail skin areas of Aspm-CreER low TM and Dlx1-CreER low TM lineage traced mice ( Table S1 ). Numbers at the top indicate the total number of clones counted.

Article Snippet: Rabbit anti-K14 (1:100, BioLegend, 905301) or mouse anti-K10 (1:100, BioLegend, 904301) and rat anti-β4-integrin (1:200, BD Biosciences) or rabbit anti-Ki67 (1:100, Abcam, ab15580) were used.

Techniques: Injection, Staining, Clone Assay

Journal: bioRxiv

Article Title: Lethal giant larvae gene family ( Llgl1 and Llgl2 ) functions as a tumor suppressor in mouse skin epidermis

doi: 10.1101/2023.03.06.531408

Figure Lengend Snippet: A . Western blot analysis of proteins from epidermises of newborn mice with indicated genotypes using anti-Llgl1 and anti-Llgl2 antibodies. B . Survival curve of Control (K14-Cre) and Llgl1/2 -/- cKO (K14-Cre/ Llgl1/2 fl/fl ) mice. Log-Rank test p=0.07. C . Confocal images of immunofluorescent staining of skin sections from Control (K14-Cre) and Llgl1/2 -/- cKO (K14-Cre/ Llgl1/2 fl/fl ) new-born pups with anti-β4 integrin (green) and anti-LLGL1/2 (red) antibodies. Areas in black boxes are shown at higher magnification on the right. DAPI (blue) indicates nuclear counter stain. Scale bar 10μm. D . Loss of hair and prominent wound like lesions in 1 year old Llgl1/2 -/- cKO individual. E-F’ . H&E staining of skin sections from K14-Cre (Ctrl) and K14-Cre/ Llgl1/2 fl/fl ( Llgl1/2 -/- cKO) mice. Images in boxes in E and F are shown at higher magnification in E’ and F’. Same magnification for E-F and E’-F’. Note prominent inflammation and lesion missing the epidermal skin layer in 1 year old Llgl1/2 -/- cKO individual (white arrows in D and F’).

Article Snippet: For Immunostaining, we used the following antibodies: rabbit anti-LLGL1 (cross-react with LLGL2 ( ) (1:100, Abcam, #183021); rat anti-Integrin β4 chain (1:100, BD Transduction Laboratories, #553745), rabbit anti-Ki67 (1:300, Novo Castra, #NCL-Ki67p). anti-mouse-conjugated Texas Red (Jackson Immunoresearch, 115-075-075) and anti-rabbit-conjugated fluorescein isothiocyanate (FITC) (Jackson Immunoresearch, 711-095-152).

Techniques: Western Blot, Staining

Journal: PLoS ONE

Article Title: Ets1 Induces Dysplastic Changes When Expressed in Terminally-Differentiating Squamous Epidermal Cells

doi: 10.1371/journal.pone.0004179

Figure Lengend Snippet: Immunofluorescent staining of epidermal differentiation markers on dorsal skin of induced Ets1 BT mice and littermate wild type mice. Each section was stained with antibodies to β4- integrin to mark the position of the basement membrane and with DAPI to detect nuclei. Epidermal differentiation was assessed by staining for (A) the basal layer marker keratin 14 (K14), (B) the spinous layer marker keratin 10 (K10) and (C) the granular layer marker loricrin (Lor). (D) Western blot analysis of dorsal skin extracts to confirm results obtained in immunofluorescence. GAPDH serves as a loading control for Western blots.

Article Snippet: The following primary antibodies were used for Western blotting, immunofluorescence and/or immunohistochemistry: rat monoclonal anti-HA (clone 3F10, Roche Applied Sciences), rabbit polyclonal anti-Ets1 (N-276, Santa Cruz Biotechnology), mouse monoclonal anti-GAPDH (clone 6C5, Chemicon International), goat polyclonal anti-MMP13 (W-16, Santa Cruz Biotechnology), mouse monoclonal anti-Ki67 (NCL-Ki67p, Novocastra), rat monoclonal anti-β4 integrin (CD104, clone 346-11A, BD Biosciences), rat monoclonal anti-PECAM1 (CD31, clone MEC-13.3, BD Biosciences), rabbit polyclonal anti-laminin I (CL54851AP, Cedarlane Laboratories) and mouse monoclonal anti-PCNA (clone PC10, Dakocytomation).

Techniques: Staining, Marker, Western Blot, Immunofluorescence

Journal: PLoS ONE

Article Title: Ets1 Induces Dysplastic Changes When Expressed in Terminally-Differentiating Squamous Epidermal Cells

doi: 10.1371/journal.pone.0004179

Figure Lengend Snippet: Gene expression profiles in WT versus BT mice.

Article Snippet: The following primary antibodies were used for Western blotting, immunofluorescence and/or immunohistochemistry: rat monoclonal anti-HA (clone 3F10, Roche Applied Sciences), rabbit polyclonal anti-Ets1 (N-276, Santa Cruz Biotechnology), mouse monoclonal anti-GAPDH (clone 6C5, Chemicon International), goat polyclonal anti-MMP13 (W-16, Santa Cruz Biotechnology), mouse monoclonal anti-Ki67 (NCL-Ki67p, Novocastra), rat monoclonal anti-β4 integrin (CD104, clone 346-11A, BD Biosciences), rat monoclonal anti-PECAM1 (CD31, clone MEC-13.3, BD Biosciences), rabbit polyclonal anti-laminin I (CL54851AP, Cedarlane Laboratories) and mouse monoclonal anti-PCNA (clone PC10, Dakocytomation).

Techniques: Expressing