Figure 5.

Ripk3 signaling induces protein synthesis and cellular senescence in HSCs in an Mlkl-independent fashion

(A–D) WT and Ripk3^−/− mice were irradiated with X-rays, 1.75 Gy weekly × 4. LK and LSK cells were collected from mouse BM 1 month after the final IR. Gene expression was examined by RNA-seq. Genes up- or downregulated in LK and LSK cells of Ripk3^−/− mice compared with WT mice are presented in the heatmaps (A and B). Signaling pathways specifically altered in the LSK and LK populations of Ripk3^−/− mice compared with WT mice were analyzed by GO enrichment analysis (C and D).

(E–I) WT, Ripk3^−/−, Mlkl^−/−, and Tnfr^−/− mice were irradiated with X-rays, 1.75 Gy weekly × 4. LK and LSK cells were collected from mouse BM 1 month after the last IR. The expression of selected genes from RNA-seq data was verified in LSK and LK cells by qRT-PCR (E). Protein synthesis (F), inflammasome activity (G), and senescence (H and I) were examined by OP-puro, a-Casp1, and C12GFDG staining, as well as p16, p19, and p15 expression (I). Data in (F), (G), and (H) show one of the three biological triplicate experiments. ^∗p < 0.01, compared with non-irradiated controls. ^&p < 0.05, compared with WT and Mlkl^−/− mice.