Validation of candidates associated with ER stress–induced autophagy. (A and B) Wild-type HEK293A cells were infected with both lentiviruses harboring both Cas9 and sgRNA targeting indicated positive (A) or negative (B) regulators. Polyclonal KO cells were then incubated with tunicamycin (10 µg/ml) for 6 h. Autophagy flux was examined through blots of p62. Experiments were repeated six times. P values denote statistical significance of treated KO cells compared with treated control cells and were determined by two-way ANOVA. *P ≤ 0.1; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001; ns, not significant.