Fig. 3: eIF2α phosphorylation and translational control drive ATF4 expression in cells exposed to mycolactone. | Cell Death & Disease

Fig. 3: eIF2α phosphorylation and translational control drive ATF4 expression in cells exposed to mycolactone.

From: Inhibition of Sec61-dependent translocation by mycolactone uncouples the integrated stress response from ER stress, driving cytotoxicity via translational activation of ATF4

Fig. 3

a Immunoblot analysis of newly synthesised puromycilated proteins prepared from HeLa cells exposed to DMSO, mycolactone (MYC) or tunicamycin (Tuni) for 12 h. Relative quantified signal intensities are shown (Mean ± SEM, n = 3 independent experiments). b HeLa-gs cells stably expressing GFP-GADD34 (clone 8) or GFP-KARA (clone 4), an inactive mutant of GADD34, were exposed for 5 h. Lysates were analysed by immunoblotting. c HeLa cells were pre-treated with ISRIB for 1 h, followed by exposure to mycolactone for 8 h. Lysates were analysed by immunoblotting. d RAW246.7 cells were exposed to either DMSO, mycolactone or 100 nM ISRIB for 5 h. For co-incubation cells were pre-treated with ISRIB for 1 h prior to addition of mycolactone. Cell lysates were subject to polysome profiling. All immunoblots show the approximate migration of molecular weight markers in kDa. All data representative of at least three independent experiments

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