Fig. 5. pMLKL-S345 staining distinguishes between RIPK3-mediated necroptosis and RIPK3-mediated inflammation.
A Representative images of mice of indicated genotypes at the endpoint (14–15 weeks). Control mice included littermates with the Shpn+/m genotype. B Kaplan–Meier survival graph of mice with the indicated genotypes (n = 7 for Shpn+/m, 6 for Shpnm/m, 1 for Shpnm/m; Ripk3−/− mice). C Representative images of skin and spleen sections from mice with the indicated genotypes stained with H&E. Slides were digitalised in a digital slide scanner and images were acquired in QuPath. D Graph of severity score of dermatitis assessment in 14–15 weeks old mice of the indicated genotypes. Shpn+/m mice (14–21 weeks old) served as controls (n = 5 for Shpn+/m, 6 for Shpnm/m, 1 for Shpnm/m; Ripk3−/− mice). E Representative bright field images of spleens from mice of the indicated genotypes. F, G Representative bright field images of skin (F) and spleen (G) sections from mice with the indicated genotypes immunostained with pMLKL-S345 and c-Casp3. Consecutive skin sections from the same mice were utilised for each marker and displayed according to their order (n = 3 for Shpn+/m, 3 for Shpnm/m and 1 for Shpnm/m; Ripk3−/− mice). Scale bars: skin (F) 100 µm (representative field) and 50 µm (magnified selected area), spleen (G) 50 µm for both representative field and magnified selected area.