Fig. 4

Ethanol-induced IL-17A release is augmented by ER stress. a Isolated small intestinal crypts were treated with ethanol (100 mM) to induce IL-17A release and some crypts were also treated with 5 or 10 mM of the ER stress inhibitor 4-phenylbutiric acid (4-PBA). Released IL-17A was measured by ELISA from the crypt supernatants. b IL17A release was measured by ELISA performed on isolated crypts treated with the ER stress inducer thapsigargin (0.3, 1, and 3 μM) or ethanol (25, 50, and 100 mM). c Cleaved (clv) PARP expression was measured by Western blot and quantified (d) from isolated crypts treated ex vivo with 10 ng/ml of recombinant IL-17A, 50 mM EtOH or IL-17A and EtOH. e Cleaved PARP, pro-caspase 8 and cleaved caspase 8 p43 subunit from the PSI of pair-fed (PF) or alcohol-fed mice treated or not with a control isotype antibody or IL17A blocking antibody were measured by Western blot and quantified (f). For a–b, n = 3 mice/treatment group; p < 0.05 * vs. untreated, # vs. EtOH 100 mM. For d–e, n = 3–4, p < 0.05 * vs. Untreated. For f–g, n = 3–4 mice/group; p < 0.05 * vs. PF