AGER mediates rSQSTM1 activity in acinar cells. (A, B) Analysis of Tnf and Il6 mRNA expression in WT and insr−/− peritoneal macrophages after treatment with rSQSTM1 (2 ng/ml) for 24 h (n = 3 biologically independent samples; two-way ANOVA with Tukey’s multiple comparisons test on all pairwise combinations; data are presented as means ± SD). (C, D) Analysis of Acsl4 mRNA expression and intracellular 5-HETE level in WT and insr−/− acinar cells after treatment with rSQSTM1 (2 ng/ml) for 24 h (n = 3 biologically independent samples; data are presented as means ± SD). (E) His-tag affinity-isolation analysis of the binding of SQSTM1 to AGER in the absence or presence of αSQSTM1. (F, G) Analysis of Acsl4 mRNA expression and intracellular 5-HETE levels in WT and ager−/− acinar cells after treatment with rSQSTM1 (2 ng/ml) for 24 h (n = 3 biologically independent samples; two-way ANOVA with Tukey’s multiple comparisons test on all pairwise combinations; data are presented as means ± SD). (H) Analysis of the level of cell death in WT and ager−/− acinar cells after treatment with cerulein (100 nM), L-arginine (5 mg/mL), erastin (5 µM), or RSL3 (500 nM) in the absence or presence of rSQSTM1 (2 ng/ml) for 48 h (n = 3 biologically independent samples; two-way ANOVA with Tukey’s multiple comparisons test on all pairwise combinations; data are presented as means ± SD).