Journal: Scientific Reports

Article Title: Descemet’s Membrane Supports Corneal Endothelial Cell Regeneration in Rabbits

doi: 10.1038/s41598-017-07557-2

Figure Lengend Snippet: Endothelial cell tracing and α-SMA staining of cornea after CEC scraping. CFDA SE-labeled cells shows the leading edge of CEC injury (arrow) on day 3 and day 7, and complete CEC healing on day 14. Immunostaining showed that CFDA SE-labeled cells close to the leading edge were strongly expressed with α-SMA on day 3 (arrow), decreased on day 7 (arrow), and became negative on day 14. Bars represent 50 μm.

Article Snippet: After 3 rinses with PBS for 5 minutes each and preincubation with 2% bovine serum albumin (BSA) for 1 hour at room temperature, sections were incubated with α-SMA antibody (1:200, ab18147, Abcam, Cambridge, MA, US) at 4 °C overnight.

Techniques: Staining, Labeling, Immunostaining

Journal: Scientific Reports

Article Title: Descemet’s Membrane Supports Corneal Endothelial Cell Regeneration in Rabbits

doi: 10.1038/s41598-017-07557-2

Figure Lengend Snippet: Endothelial cell tracing and α-SMA staining of the cornea after DM stripping. CFDA SE-labeled cells showed minor migration toward the central cornea from day 3 to day 14. Immunostaining showed that some CFDA SE-labeled cells close to the wound edge (arrows) expressed α-SMA on day 3. On day 7, cells on the stromal surface of the surgical site showed positive α-SMA staining, but they were not labeled with CFDA SE. CFDA SE-labeled α-SMA positive cells became negative on day 7. Cells on the stromal surface of the surgical site remained positive α-SMA staining on day 7 and day 14. Bars represent 50 μm.

Article Snippet: After 3 rinses with PBS for 5 minutes each and preincubation with 2% bovine serum albumin (BSA) for 1 hour at room temperature, sections were incubated with α-SMA antibody (1:200, ab18147, Abcam, Cambridge, MA, US) at 4 °C overnight.

Techniques: Staining, Stripping Membranes, Labeling, Migration, Immunostaining

Journal: Scientific Reports

Article Title: Descemet’s Membrane Supports Corneal Endothelial Cell Regeneration in Rabbits

doi: 10.1038/s41598-017-07557-2

Figure Lengend Snippet: Whole-mount staining of ZO-1 and α-SMA on a CEC scraping cornea. ZO-1 expression decreased in the forefront of the migrating cells, while α-SMA was expressed in the migrating CECs on day 7. At day 14, ZO-1 was re-expressed on the cell border, although it was not as organized as that in the normal endothelium, while α-SMA became negative in all the endothelial cells. Bars represent 20 μm.

Article Snippet: After 3 rinses with PBS for 5 minutes each and preincubation with 2% bovine serum albumin (BSA) for 1 hour at room temperature, sections were incubated with α-SMA antibody (1:200, ab18147, Abcam, Cambridge, MA, US) at 4 °C overnight.

Techniques: Staining, Expressing

Journal: Scientific Reports

Article Title: Descemet’s Membrane Supports Corneal Endothelial Cell Regeneration in Rabbits

doi: 10.1038/s41598-017-07557-2

Figure Lengend Snippet: Whole-mount staining of ZO-1 and α-SMA on a DM stripping cornea. ZO-1 expression in the endothelial cells close to the wound border decreased on day 7, cells on the stromal surface of the wound area were not labeled with CFDA SE but were highly expressed with α-SMA. On day 14, ZO-1 expression was regained in the endothelial cells close to the wound border. CFDA SE negative cells close to the wound border also showed positive ZO-1 expression. α-SMA was maintained in the CFDA SE negative cells, while it was negative in the CFDA SE-labeled cells. Bars represent 20 μm.

Article Snippet: After 3 rinses with PBS for 5 minutes each and preincubation with 2% bovine serum albumin (BSA) for 1 hour at room temperature, sections were incubated with α-SMA antibody (1:200, ab18147, Abcam, Cambridge, MA, US) at 4 °C overnight.

Techniques: Staining, Stripping Membranes, Expressing, Labeling

Journal: Scientific Reports

Article Title: Descemet’s Membrane Supports Corneal Endothelial Cell Regeneration in Rabbits

doi: 10.1038/s41598-017-07557-2

Figure Lengend Snippet: Gene expression in corneal endothelial cells and stromal cells after endothelial injury. ZO-1 gene expression in CECs after scraping injury was significantly higher than that in CECs after DM stripping at day 3, 7, and 14 ( A ). Na + /K + ATPase gene expression significantly decreased after DM stripping at day 3, 7, and 14 ( B ). α-SMA gene expression in CECs dramatically increased after scraping injury and DM stripping at day 3, 7, and 14. α-SMA expression was higher in CEC scraping group at day 3, while was higher in DM stripping group at day 7 and day 14 ( C ). α-SMA gene expression in central corneal stroma showed mild increase in CEC scraping group at day 3, 7, and 14. While there was dramatic increase in DM stripping group at day 7 and day 14 ( D ). (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: After 3 rinses with PBS for 5 minutes each and preincubation with 2% bovine serum albumin (BSA) for 1 hour at room temperature, sections were incubated with α-SMA antibody (1:200, ab18147, Abcam, Cambridge, MA, US) at 4 °C overnight.

Techniques: Expressing, Stripping Membranes