Journal: bioRxiv

Article Title: KRT6A and KRT17 mark distinct stem cell populations in the adult palpebral conjunctiva and Meibomian gland

doi: 10.1101/2025.06.10.658923

Figure Lengend Snippet: (A) Schematic depicting the breeding strategy and experimental setup. (B, C) IF results demonstrate that Krt17 GFP + cells are absent in PC epithelium (blue arrow, ), but are present in MG duct suprabasal basal layer (red arrow, and ), duct basal layer (white arrow, ), acinar basal layer (pink arrow, ), and ductule (yellow arrow, ) in mice following 2 days of lineage tracing. (D, E) IF identifies Krt17 GFP + cells in MG duct (white arrows in and ) and acini (pink arrows in and ) in mice following 3 months (D) or 18 months (E) of lineage tracing. The yellow dashed line in (B) delineates PC, while the white dashed lines in (C-E) delineate the MG duct. KRT14 marks epithelial cells. Scale bars: 25 μm.

Article Snippet: The following primary antibodies were used: rabbit anti-HDAC3 (Santa Cruz, sc-11417, 1:100); Guinea pig anti-PLIN2 (Fitzgerald, 20R-AP002, 1:500); rabbit anti-GFP (Abcam, ab290, 1:1000), rabbit anti-KRT6A (BioLegend, 905702, 1:200); mouse anti-BrdU (Cell signaling, 5292S, 1:200), mouse anti-MYC-tag (Cell Signaling, 2276S, 1:1000); goat anti-KLF4 (R&D Systems, AF3158, 1:100); chicken anti-KRT14 (BioLegend, 906004, 1:500); rat anti-Ki-67 (eBioscience, 14-5698-82, 1:200); rabbit anti-KRT17 (Abcam, ab109725, 1:200).

Techniques:

Journal: bioRxiv

Article Title: Topical Delivery of 4-Aminopyridine Enhances Skin Regeneration in Burn Wounds

doi: 10.1101/2025.06.05.658142

Figure Lengend Snippet: B) Representative IF images and quantitative results of keratinocyte and hair follicle regeneration markers (K14 and K15) in skin burn mice treated with control or 4-AP gels on day 21. Scale bar = 100 µm. n = 6 mouse skin tissue samples per group. Protein (C and D) and gene expression results (E) indicate that the 4-AP gel enhances the keratin markers K14 and K15 on day 21 post-skin burn injury, compared to the control group. Data from three skin tissue samples per group are shown as mean ± SEM, with significance marked (*P < 0.05 and **P < 0.0021). Comparisons were performed using two-tailed, unpaired t-tests.

Article Snippet: The following antibodies were applied by incubating the slides overnight at 4 °C: K14 (1:100, # NBP2-34270, Novus Biologicals), K15 (1:500, #833901, BioLegend), Vimentin (1:200, # 10366-1-AP, Thermo Fisher Scientific), α-SMA (1:500, #14-9760-82, Thermo Fisher Scientific), and TGF-β (1:500, # 3711s, Cell Signaling).

Techniques: Control, Gene Expression, Two Tailed Test

Journal: Investigative Ophthalmology & Visual Science

Article Title: SLC6A6-Mediated Taurine Uptake Sustains Corneal Epithelial Stem/Progenitor Cell Function to Counteract Age-Related Dysfunction

doi: 10.1167/iovs.66.6.25

Figure Lengend Snippet: Slc6a6 was predominantly expressed in corneal epithelial stem/progenitor cells. ( A ) UMAP plots of corneal epithelial cell subpopulations. LSC, limbal stem cells; TAC1/2, transient amplifying cells 1/2; Basal, basal cells; Wing, wing cells; Squam: squamous cells. ( B ) Dot plot of marker gene expression patterns. ( C ) Monocle pseudotime trajectory analysis colored by cell clusters ( left panel ) and feature plots of Krt14 , Muc4 , and Slc6a6 across pseudotime ( right panel ). ( D ) Violin plots of Krt14 and Slc6a6 expression in corneal epithelial cells. ( E ) The expression of Slc6a6 in the corneal epithelium of C57BL/6J mouse and human donor corneas.

Article Snippet: Corneas were permeabilized and blocked overnight at 4°C in blocking buffer (PBS containing 0.3% Triton X-100 and 5% bovine serum albumin [BSA]), and then incubated with anti-SLC6A6 (26725-1-AP, Proteintech, Chicago, IL, USA) or anti-KRT14 (IY1383542, Invitrogen, Carlsbad, CA, USA) antibodies at 4°C overnight.

Techniques: Marker, Gene Expression, Expressing

Journal: Investigative Ophthalmology & Visual Science

Article Title: SLC6A6-Mediated Taurine Uptake Sustains Corneal Epithelial Stem/Progenitor Cell Function to Counteract Age-Related Dysfunction

doi: 10.1167/iovs.66.6.25

Figure Lengend Snippet: SLC6A6 decreased in the corneal epithelial stem/progenitor cells of aged mice. ( A ) UMAP plots of corneal epithelial cells from young and aged mice. ( B ) Violin plots of Krt14 and Slc6a6 expression in stem/progenitor cells. ( C ) The expression of SLC6A6 in the corneal epithelium of young and aged mice ( left panel , n = 4), and the fluorescence intensity was quantified by ImageJ ( right panel ). ( D , E ) Gene scoring analysis using genes related to stemness ( D ) and differentiation ( E ). ( F ) The corneal epithelium of young mice (3 months old, female) and aged mice (11 months old, female) was stained with fluorescein sodium at 0, 24, 48, and 60 hours ( left panel , n = 7) after wounding, and the corneal wound closure was presented as the percentage of the original wound ( right panel ). Data are shown as mean ± SD. ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: Corneas were permeabilized and blocked overnight at 4°C in blocking buffer (PBS containing 0.3% Triton X-100 and 5% bovine serum albumin [BSA]), and then incubated with anti-SLC6A6 (26725-1-AP, Proteintech, Chicago, IL, USA) or anti-KRT14 (IY1383542, Invitrogen, Carlsbad, CA, USA) antibodies at 4°C overnight.

Techniques: Expressing, Fluorescence, Staining

Journal: Investigative Ophthalmology & Visual Science

Article Title: SLC6A6-Mediated Taurine Uptake Sustains Corneal Epithelial Stem/Progenitor Cell Function to Counteract Age-Related Dysfunction

doi: 10.1167/iovs.66.6.25

Figure Lengend Snippet: Slc6a6 was predominantly expressed in corneal epithelial stem/progenitor cells. ( A ) UMAP plots of corneal epithelial cell subpopulations. LSC, limbal stem cells; TAC1/2, transient amplifying cells 1/2; Basal, basal cells; Wing, wing cells; Squam: squamous cells. ( B ) Dot plot of marker gene expression patterns. ( C ) Monocle pseudotime trajectory analysis colored by cell clusters ( left panel ) and feature plots of Krt14 , Muc4 , and Slc6a6 across pseudotime ( right panel ). ( D ) Violin plots of Krt14 and Slc6a6 expression in corneal epithelial cells. ( E ) The expression of Slc6a6 in the corneal epithelium of C57BL/6J mouse and human donor corneas.

Article Snippet: The PVDF membranes were blocked by nonfat dry milk for 2 hours at room temperature and incubated with anti-GAPDH (60004-1-Ig; Proteintech), anti-KRT14, anti-p63, anti-BCAM, anti-β-actin (66009-1-Ig; Proteintech), or anti-Notch1 antibody in universal antibody diluent (Epizyme Biomedical Technology) overnight at 4°C.

Techniques: Marker, Gene Expression, Expressing

Journal: Investigative Ophthalmology & Visual Science

Article Title: SLC6A6-Mediated Taurine Uptake Sustains Corneal Epithelial Stem/Progenitor Cell Function to Counteract Age-Related Dysfunction

doi: 10.1167/iovs.66.6.25

Figure Lengend Snippet: SLC6A6 decreased in the corneal epithelial stem/progenitor cells of aged mice. ( A ) UMAP plots of corneal epithelial cells from young and aged mice. ( B ) Violin plots of Krt14 and Slc6a6 expression in stem/progenitor cells. ( C ) The expression of SLC6A6 in the corneal epithelium of young and aged mice ( left panel , n = 4), and the fluorescence intensity was quantified by ImageJ ( right panel ). ( D , E ) Gene scoring analysis using genes related to stemness ( D ) and differentiation ( E ). ( F ) The corneal epithelium of young mice (3 months old, female) and aged mice (11 months old, female) was stained with fluorescein sodium at 0, 24, 48, and 60 hours ( left panel , n = 7) after wounding, and the corneal wound closure was presented as the percentage of the original wound ( right panel ). Data are shown as mean ± SD. ** P < 0.01, *** P < 0.001, **** P < 0.0001.

Article Snippet: The PVDF membranes were blocked by nonfat dry milk for 2 hours at room temperature and incubated with anti-GAPDH (60004-1-Ig; Proteintech), anti-KRT14, anti-p63, anti-BCAM, anti-β-actin (66009-1-Ig; Proteintech), or anti-Notch1 antibody in universal antibody diluent (Epizyme Biomedical Technology) overnight at 4°C.

Techniques: Expressing, Fluorescence, Staining