Fig. 3: GDF15 limits alcohol-associated liver inflammation by inducing apoptosis of perivenous KCs.

a–f GDF15^f/f and GDF15^AlbCre mice were fed EtOH for 8 weeks (n = 5/group; 3 biological replicates). a Portal catecholamine levels were measured. b Serum ALT and GDF15 levels were measured. c Representative immunostaining (left) and the number of CLEC4F⁺ KCs in the CYP2E1⁺ area in the liver (right). d Annexin V apoptosis assay of isolated KCs. e Flow cytometry analysis of F4/80^lowCD11b⁺ hepatic macrophages with representative plots. f qRT‒PCR analyses of isolated KCs from EtOH-fed GDF15^f/f and GDF15^AlbCre mice (n = 3/group). g WT KCs were pretreated with LPS (100 ng ml⁻¹, 3 h) followed by the indicated doses of GDF15 for 6 h. qRT‒PCR for the mRNA levels of adrenergic receptors (3 replicates). h WT KCs were pretreated with LPS (100 ng ml⁻¹) or KT5720 (PKA inhibitor, 1 μM) for 6 h, followed by GDF15 (100 ng ml⁻¹) or CBL (1 μM) treatment for 6 h (3 replicates). Immunoblots for ADRB2, PKA-Cα, Bcl-2, cleaved-caspase 9, and cleaved-caspase 3 (left) and Annexin V assay of KCs (right). Data are presented as the mean ± SEM. *p < 0.05, **,^##p < 0.01, ***p < 0.001 by Student’s t test or by one-way ANOVA. Scale bars, 50 µm.