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. 2022 Oct 26;23(21):12983. doi: 10.3390/ijms232112983

Figure 4.

Figure 4

Effect of valdecoxib on the expression of ER stress-related and apoptosis-related proteins in the retinal ischemia-reperfusion model. (A) Western blot analysis of p-PERK, ATF4, GRP78, CHOP, cleaved caspase 3, bax and bcl-2 levels in the control, I/R, I/R+DMSO and I/R+VAL groups. β-tubulin served as the loading control. (B–H) Quantification of expression levels of p-PERK, ATF4, GRP78, CHOP, cleaved caspase 3, bax and bcl-2 in the control, I/R, I/R+DMSO and I/R+VAL groups using the densitometric analyses of Western blotting. The bar charts show the quantitative data (normalized by β-tubulin) for each protein relative to the sham group (assigned a value of 1). (I) Representative fluorescence images of p-PERK staining are shown (scale bar = 50 μm). Immunostaining was executed using a primary antibody against p-PERK (green), and the nucleus (blue) is marked by DAPI. (J) Representative fluorescence images of cleaved caspase 3 staining are shown (scale bar = 50 μm). Immunostaining was executed using a primary antibody against cleaved caspase 3 (green), and the nucleus (blue) is marked by DAPI. (K) Representative fluorescence images of GRP78 staining are shown (scale bar = 50 μm). Immunostaining was executed using a primary antibody against GRP78 (green), and the nucleus (blue) is marked by DAPI. Images demonstrate the increased expression level of p-PERK, cleaved caspase 3 and GRP78 in the I/R and I/R+DMSO groups, and decreased expression level of those proteins in the I/R+VAL group in the retinal ganglion cell layer. Data are represented as the mean ± SD of three independent experiments. Each group was composed of five rats. One-way ANOVA is used in B to H. * p < 0.05, ** p < 0.01, *** p < 0.001 vs. sham group. ## p < 0.01, ### p < 0.001 vs. I/R group. $$ p < 0.01, $$$ p < 0.001 vs. I/R+DMSO group.