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. 2022 Apr 29;298(6):101997. doi: 10.1016/j.jbc.2022.101997

Figure 4.

Figure 4

Effects of S724A mutation upon cell survival under ER stress in MEF cells. MEF cells of the indicated genotypes were treated with 10 μg/ml Tm for the indicated time intervals. A, immunoblot analysis of PERK phosphorylation and CHOP protein levels. Tubulin was used as the loading control. B, immunofluorescence staining of CHOP protein along with DAPI staining in MEF cells following Tm treatment. The scale bar represents 50 μm. C, cell viability of Tm-treated MEF cells was determined by CCK8 assay. Shown is the percentage of viability after normalization to their untreated controls. D, immunoblot analysis of cleavage of caspase-3 protein. Tubulin was used as the loading control. Data are presented as the mean ± SD (n = 3 independent experiments). ∗∗∗p < 0.001 by two-tailed unpaired Student’s t test. CCK8, Cell Counting Kit-8; CHOP, CCAAT-enhancer binding protein homologous protein; DAPI, 4′,6-diamidino-2-phenylindole; ER, endoplasmic reticulum; MEF, mouse embryonic fibroblast; PERK, PKR-like endoplasmic reticulum kinase; Tm, tunicamycin.