Journal: Biomolecules
Article Title: Effect of Topical Programmed Death-Ligand1 on Corneal Epithelium in Dry Eye Mouse.
doi: 10.3390/biom14010068
Figure Lengend Snippet: Figure 3. (A) Immunofluorescence staining showing the expression of (i) PD-L1, (ii) CD4, (iii) CD3e and (iv) IL-17 localized in the corneas in the control, desiccation+PBS and desiccation+PD-L1 group. (B–E) Changes in the percentage of PD-L1, CD4, CD3e and IL-17 positive cells in the cornea. Im- munopositivity for PD-L1 is shown in higher-power fields. Immunopositivity was counted in low-power fields and calculated as relative to the total number of DAPI-positive cells. (F,G) Western blot showing the expression of CD4 and IL-17 in the cornea. (H,I) Relative expression ratio of CD4 and IL-17 to β-actin. In (B–E,H,I), data are presented with bar graph (n = 6, day 10, * p < 0.05; ** p < 0.01; *** p < 0.001 significant difference by one-way ANOVA). Original images of (F,G) can be found in Supplementary Materials.
Article Snippet: The membranes were blocked by incubation with 5% BSA in Trisbuffered saline containing Tween 20 TBST for 1 h at room temperature and incubated overnight at 4 ◦C with specific rabbit polyclonal antibodies against CD4 (1:1000, Catalog No. NBP1-19371; Novus Biologicals, Littleton, CO, USA), IL-17 (1:1000, Catalog No. ab79056; Abcam, Inc.), phosphorylated NF-kB (pNF-kB; 1:1000, Catalog No. 3033S; Cell Signaling Technology, Inc.), NF-kB (1:1000, Catalog No. 51-0500; Invitrogen, Inc.), phosphorylated IkB-α (pIkB-a 1:1000, Catalog No. 9246S; Cell Signaling Technology, Inc.), IkB-α (1:1000, Catalog No. PA5-17888; Invitrogen, Inc.), Bax protein (1:1000, Catalog No. sc-20067; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) and β-actin (1:10,000, Catalog No. 5125S; Cell Signaling Technology, Inc.).
Techniques: Immunofluorescence, Staining, Expressing, Control, Western Blot