Fig. 4: Necroptosis triggers galectin-1 release.

a, Galectin-1 and LDH release by L929 cells stimulated with DMSO, TNF + zVAD and poly(I:C) + zVAD in the presence or absence of the RIPK3 inhibitors GSK872 or GSK843 (1 μM). Cells were treated with inhibitors 1 h before stimulation and supernatants were collected at 18 h. The immunoblot of phospho-MLKL and total MLKL in the lysates of L929 cells stimulated as above is shown. b, Galectin-1 and LDH release by RAW 264.7 macrophages stimulated with DMSO, LPS + zVAD, poly(I:C) + zVAD and the SMAC mimetic LCL-161 + zVAD in the presence or absence of the RIPK3 inhibitors GSK872 or GSK843 (2.5 μM). Cells were treated with inhibitors 1 h before stimulations and supernatants were collected at 18 h. Immunoblot of phospho-MLKL and total MLKL in the lysates of RAW 264.7 macrophages stimulated as above. c, Galectin-1 levels in the plasma (left) and peritoneal cavity (right) of WT mice (n = 6) injected intraperitoneally with 500 μg kg⁻¹ TNF at 6, 12 and 20 h. Combined data from three independent experiments are shown as the mean ± s.e.m. a,b, Immunoblots are representative of two independent experiments. c, Combined data from two independent experiments are shown; each circle represents a mouse and the horizontal lines represent the mean (n = 6). *P < 0.05; two-way (a,b) or oneway (c) ANOVA followed by Šidák’s post-test.