Journal: Scientific Reports

Article Title: Caveolin 1 and 2 enhance the proliferative capacity of BCAM-positive corneal progenitors

doi: 10.1038/s41598-024-81283-4

Figure Lengend Snippet: Evaluation of CAV1 and CAV2 expression in human limbus and cornea. Comparative analyses of CAV1 and CAV2 mRNA expression in BCAM-positive and BCAM-negative cells in the limbus ( a ) ( n = 8) and cornea ( b ) ( n = 3) (* p < 0.05, ** p < 0.01) ( c ) Representative immunostaining analyses of CAV1 (green), CAV2 (yellow) and BCAM (red) co-expression in the limbus and cornea. Sequential sections were used to illustrate BCAM and CAV1 co-expression since both antibodies were raised in rabbits. Nuclei are visualized with Hoechst 33342 (blue). n = 3. Scale bar, 50 μm. ( d ) Representative immunostaining analyses of CAV1 (green), CAV2 (yellow) and Laminin α5 (red) co-expression (top panels) and CAV1 (green), CAV2 (yellow) and Laminin α3 (red) (bottom panels) in the limbus and the cornea. Nuclei are visualized with Hoechst 33342 (blue). n = 3. Scale bar, 50 μm.

Article Snippet: The following primary antibodies were used: mouse anti-CAV1 mAb (1:100, Santa Cruz Biotechnology, Santa Cruz, CA) for co-staining with FGFR2, rabbit anti-CAV1 pAb (1:200, GeneTex, Irvine, CA) for co-staining with CAV2, Laminin α5 and Laminin α3, goat anti-CAV2 pAb (1:100, R&D Systems, Minneapolis, MN), rabbit anti-BCAM polyclonal antibody (pAb) (1:100, Novus Biologicals, Centennial, CO), mouse anti-Laminin α5 mAb (1:50, Atlas Antibodies, Bromma, Sweden), mouse anti-Laminin α3 mAb (1:100, Atlas Antibodies), rabbit anti-FGFR2 mAb (1:100, Cell Signaling Technology, Danvers, MA).

Techniques: Expressing, Immunostaining

Journal: Scientific Reports

Article Title: Caveolin 1 and 2 enhance the proliferative capacity of BCAM-positive corneal progenitors

doi: 10.1038/s41598-024-81283-4

Figure Lengend Snippet: Regulation of CAV1 and CAV2 expression in limbal epithelial cells. ( a ) Left, western blot analysis of CAV1, CAV2 and BCAM expression in BCAM KD limbal epithelial cells. Right, bar graph depicts the quantitative analyses of CAV1 and CAV2 protein expression. Bottom, representative flow cytometry analysis of BCAM expression in BCAM KD limbal epithelial cells. n = 4, * p < 0.05, KD, knockdown. ( b ) Left, western blot analyses of CAV1, CAV2 and laminin α5 expression in LAMA5 KD limbal epithelial cells. Right, the bar graphs represent the quantitative analyses of CAV1 and CAV2 protein expression. n = 8, *** p < 0.001, **** p < 0.0001. ( c ) Left, Western blot analyses of CAV1, CAV2 and laminin α3 expression in control and LAMA3 KD limbal epithelial cells. Right, the bar graphs represent the quantitative analyses of CAV1 and CAV2 protein expression. n = 6, * p < 0.05, ** p < 0.01.

Article Snippet: The following primary antibodies were used: mouse anti-CAV1 mAb (1:100, Santa Cruz Biotechnology, Santa Cruz, CA) for co-staining with FGFR2, rabbit anti-CAV1 pAb (1:200, GeneTex, Irvine, CA) for co-staining with CAV2, Laminin α5 and Laminin α3, goat anti-CAV2 pAb (1:100, R&D Systems, Minneapolis, MN), rabbit anti-BCAM polyclonal antibody (pAb) (1:100, Novus Biologicals, Centennial, CO), mouse anti-Laminin α5 mAb (1:50, Atlas Antibodies, Bromma, Sweden), mouse anti-Laminin α3 mAb (1:100, Atlas Antibodies), rabbit anti-FGFR2 mAb (1:100, Cell Signaling Technology, Danvers, MA).

Techniques: Expressing, Western Blot, Flow Cytometry, Knockdown, Control

Journal: Scientific Reports

Article Title: Caveolin 1 and 2 enhance the proliferative capacity of BCAM-positive corneal progenitors

doi: 10.1038/s41598-024-81283-4

Figure Lengend Snippet: Contribution of CAV1 and CAV2 to colony-forming efficiency. ( a ) Left, western blot analyses of CAV1 expression in CAV1 and CAV2 KD limbal epithelial cells. Right, the bar graph depicts quantitative analyses of CAV1 protein expression. n = 9. * p < 0.05, **** p < 0.0001, KD, knockdown. ( b ) Left, western blot analyses of CAV2 expression in CAV1 and CAV2 KD limbal epithelial cells. Right, the bar graph illustrates quantitative analyses of CAV2 protein expression. n = 9. ** p < 0.01, **** p < 0.0001. ( c ) Left, representative macroscopic images of the colonies formed by CAV1 and CAV2 KD cells compared to the control siRNA transfected cells. Right, the bar graph represents comparative analyses of colony-forming efficiency (CFE). n = 7. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: The following primary antibodies were used: mouse anti-CAV1 mAb (1:100, Santa Cruz Biotechnology, Santa Cruz, CA) for co-staining with FGFR2, rabbit anti-CAV1 pAb (1:200, GeneTex, Irvine, CA) for co-staining with CAV2, Laminin α5 and Laminin α3, goat anti-CAV2 pAb (1:100, R&D Systems, Minneapolis, MN), rabbit anti-BCAM polyclonal antibody (pAb) (1:100, Novus Biologicals, Centennial, CO), mouse anti-Laminin α5 mAb (1:50, Atlas Antibodies, Bromma, Sweden), mouse anti-Laminin α3 mAb (1:100, Atlas Antibodies), rabbit anti-FGFR2 mAb (1:100, Cell Signaling Technology, Danvers, MA).

Techniques: Western Blot, Expressing, Knockdown, Control, Transfection

Journal: Scientific Reports

Article Title: Caveolin 1 and 2 enhance the proliferative capacity of BCAM-positive corneal progenitors

doi: 10.1038/s41598-024-81283-4

Figure Lengend Snippet: Maintenance of cell surface FGFR2 expression by CAV1 and CAV2. (a ) Representative immunostaining analyses of CAV1 (green), CAV2 (yellow) and FGFR2 (red) expression in the limbus and cornea. Nuclei are visualized with Hoechst 33342 (blue). n = 3. Scale bar, 50 μm. ( b ) Gene expression of FGFR2 in control and CAV1 and CAV2 siRNA-treated limbal epithelial cells. n = 5. * p < 0.05, ** p < 0.01, KD, knockdown. ( c ) Left, western blot analyses of FGFR2 expression in CAV1 and CAV2 KD limbal epithelial cells. Right, bar graph illustrates quantitative analyses of FGFR2 protein expression. n = 4. * p < 0.05. ( d ) Left, representative flow cytometry analyses of FGFR2 expression in CAV1 and CAV2 KD limbal epithelial cells. Right, the bar graphs represent the quantitative analyses of FGFR2 expression. n = 5, * p < 0.05, ** p < 0.01, KD, knockdown, FSC, forward scatter; A, area.

Article Snippet: The following primary antibodies were used: mouse anti-CAV1 mAb (1:100, Santa Cruz Biotechnology, Santa Cruz, CA) for co-staining with FGFR2, rabbit anti-CAV1 pAb (1:200, GeneTex, Irvine, CA) for co-staining with CAV2, Laminin α5 and Laminin α3, goat anti-CAV2 pAb (1:100, R&D Systems, Minneapolis, MN), rabbit anti-BCAM polyclonal antibody (pAb) (1:100, Novus Biologicals, Centennial, CO), mouse anti-Laminin α5 mAb (1:50, Atlas Antibodies, Bromma, Sweden), mouse anti-Laminin α3 mAb (1:100, Atlas Antibodies), rabbit anti-FGFR2 mAb (1:100, Cell Signaling Technology, Danvers, MA).

Techniques: Expressing, Immunostaining, Gene Expression, Control, Knockdown, Western Blot, Flow Cytometry