1. ER stress response and susceptibility to apoptosis are regulated by TFEB and TFE3
ER stress response and susceptibility to apoptosis are regulated by TFEB and TFE3 AImmunoblots of protein lysates from Null‐ or TFEB/3 knockout‐MEF cells incubated in the presence of DMSO (Control) or tunicamycin (Tun.) (0.1 μg/ml) for the indicated times. Data are representative of three independent experiments.BQuantification of ATF4 protein levels in Null‐ or TFEB/3 knockout‐MEF cells treated as indicated in (A) (mean ± SD of the ATF4 to actin protein ratio from three independent experiments, Student's t‐test versus the corresponding time point in Null‐MEF cells, ns = not significant, **P < 0.01).CCell viability was determined in Null‐ or TFEB/3 knockout‐MEF cells incubated in the presence of DMSO (Control) or tunicamycin (Tun.) (0.1 μg/ml) for the indicated times (mean ± SD of the percentage of viable cells compared to control from three independent experiments, one‐way ANOVA versus the corresponding time point in Null‐MEF cells, ***P < 0.001).DChromatin condensation, an indication of cells undergoing apoptosis, was assessed in Null‐ or TFEB/3 knockout‐MEF cells incubated in the presence of DMSO (Control) or tunicamycin (0.1 μg/ml) for the indicated times. Confocal microscopy images were obtained from fixed cells stained with Hoechst Scale bar, 20 μm. Data are representative of three independent experiments.EImmunoblots of protein lysates from Null‐ or TFEB/3 knockout‐MEF cells treated as described in (A). Data are representative of three independent experiments.F–HQuantification of CHOP (F), PUMA (G), and NOXA (H) protein levels in Null‐ or TFEB/3 knockout‐MEF cells treated as indicated in (E) (mean ± SD of the indicated protein to actin ratio from three independent experiments, Student's t‐test analysis versus the corresponding time point in Null‐MEF cells, ns = not significant, *P < 0.05, **P < 0.01).IImmunoblots of protein lysates from ARPE‐19 cells infected with either adenovirus expressing Null, TFEB‐S211A, or TFE3. Data are representative of three independent experiments.JQuantification of PUMA protein levels in ARPE‐19 cells infected as indicated in (I) (mean ± SD of the fold change of the indicated protein to actin ratio from three independent experiments, one‐way ANOVA versus the adenovirus‐Null‐infected cells, *P < 0.05, ***P < 0.001). Source data are available online for this figure.
José A Martina et al. EMBO J. 2016;35:
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