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. 2023 Dec 25;13(1):42. doi: 10.3390/antiox13010042

Figure 7.

Figure 7

Inhibition of HSPA8 alleviates OGD-induced fibroblasts senescence. (A) Subcellular fractionation of NIH3T3 cells treated with OGD for 4 h were immunoprecipitated using HSPA8 antibody, and bound proteins were analyzed using Western blot. (B) Changes in ALDH2 activity in NIH3T3 cells with the siHSPA8 transfection. *** p < 0.001 compared with the Con + Nc group. # p < 0.05, compared with the OGD + Nc group. (C) β-GAL activity analysis in NIH3T3 cells with the siHSPA8 transfection. *** p < 0.001compared with the Con + Nc group, ## p < 0.01 compared with the OGD + Nc group. (D) β-GAL staining in NIH3T3 cells with the siHSPA8 transfection. (E) Western blot analysis of p53, p21, and p16 in NIH3T3 cells. (F) Quantification of (E). * p < 0.05, ** p < 0.01 compared with the Con + Nc group. # p < 0.05 compared with the OGD + Nc group. (G) RT-qPCR analysis of IL6, HMGB1, and TNF-α mRNA in NIH3T3 cells. ** p < 0.01, *** p < 0.001 compared with the Con + Nc group. # p < 0.05, ## p < 0.01 compared with the OGD + Nc group. (H) Cell proliferation was detected using Ki67 staining. The red fluorescence in the image indicates the Ki67 positive cells, while the blue fluorescence indicates the nucleus. Scale bar = 50 μm. (I) Cell lysates were treated with disuccinimidyl suberate (DSS) for 30 min and protein levels of ALDH2 were examined. (J) Quantification of (I). ** p < 0.01 compared with the CON group, # p < 0.05 compared with the OGD group. Nc: negative control.