Fig. 4: CHOP is required for ATF4-induced IOP elevation.
From: ATF4 leads to glaucoma by promoting protein synthesis and ER client protein load
a GTM3 cells were transduced with Ad5.Empty, ATF4, and CHOP for 24 h. Cellular lysates were subjected to Western blot analysis of ATF4 and CHOP. Increased CHOP expression levels were observed in ATF4-transduced TM cells whereas CHOP transduction did not alter ATF4 levels in TM cells (n = 3 replicates). b GTM3 cells transduced with Ad5.Empty, ATF4, and CHOP for 24 h. Cellular lysates were immunoprecipitated using CHOP antibody and immunoblotted for ATF4. IgG antibody was used as a negative control (n = 3 replicates). Experiments were replicated three times independently and similar results were observed in NTM-5 cell line. c 3-month-old WT (CHOP+/+) and CHOP knockout (CHOP−/−) mice were intravitreally injected with Ad5.Empty or ATF4. A significant IOP elevation was observed in CHOP+/+ mice injected with Ad5.ATF4 (n = 8) compared to CHOP+/+ mice injected with Ad5.Empty (n = 6). However, ATF4 did not significantly elevate IOP in CHOP−/− mice (n = 8) compared to Ad5. Empty injected WT mice (data are presented as mean ± SEM, two-way ANOVA with Bonferroni’s multiple comparison test, *P < 0.05, ***P < 0.001).
