Extended Data Fig. 7: Treatment with EGLNs inhibitors promotes RIPK1 activation.
(a-e) TUNEL assays (n = 5) (a), immunostaining for CC3 (n = 5) (b), serum ALT and AST levels (n = 7) (c), serum Bilirubin levels (n = 7) (d) from Ripk1P196A/P196A and control mice. DAPI for nuclei. Scale bar, 100 μm. Microscopic quantification of TUNEL, CC3 positive cells (right). Data are means ± s.e.m., Unpaired two-tailed t-test. qRT-PCR analysis of the mRNA expression of the indicated cytokines and chemokines from Ripk1P196A/P196A and control mice (n = 10) or Ripk1P196A/P196A treated with Nec-1s (n = 5) for 7 days. Data are means ± s.e.m., One-way ANOVA, post hoc Dunnett’s test (e). (f) A schematic representation of FG-4592 induction of RIPK1 activation and inflammation. (g-h) qRT-PCR analysis of the mRNA expression of HIF pathway downstream targets Glut1 and Vegf-a from livers (n = 6) (g), TUNEL assays and immunostaining for p-RIPK1(S166) on intestine sections (n = 5) (h) of 16-20-week-old mice with the indicated genotypes treated with or without FG-4592 at the dose of 25 mg/kg body weight for 7 days. Data are means ± s.e.m., One-way ANOVA, post hoc Dunnett’s test. DAPI for nuclei. Microscopic quantification of TUNEL and p-RIPK1(S166) positive cells (bottom). (i-l) Immunostaining for p-RIPK1(S166) (i), TUNEL assays (j), Serum ALT and AST levels (k), ELISA analyses of CCL2 and IL6 protein expression (l) from Ripk1P196A/P196A and control mice treated with or without FG-4592 at the dose of 250 mg/kg body weight for 2 days (n = 8). DAPI for nuclei. Scale bar, 100 μm. Microscopic quantification of p-RIPK1(S166) and TUNEL positive cells (right). Data are means ± s.e.m., Unpaired two-tailed t-test.
