Figure 2
In vitro and in vivo expression of RIP3-WT and RIP3 mutants in DA1-3b cells. (a) Western blot analysis of GFP, RIP3-WT, RIP3-KD, RIP3-RHIM expression in DA1-3b cells 10 h after the addition of 1 mM IPTG. (b) The expression of RIP3-WT and RIP3 mutants in DA1-3b cells via flow cytometry 10 h after the addition of 1 mM IPTG. (c) Survival of mice injected intraperitoneally with 1 × 106 DA1-3b/RIP3-WT and DA1-3b/RIP3-KD cells (10 mice/group). IPTG (12 mM) was added to the drinking water of the mice at day 10 (P<0.01, Log-rank test). (d) DNA fragmentation in DA1-3b/GFP, DA1-3b/RIP3-WT, DA1-3b/RIP3-KD, and DA1-3b/RIP3-RHIM cells 10 h after the addition of 1 mM IPTG. (e) Electron microscopy was performed on DA1-3b/GFP, DA1-3b/RIP3-WT, DA1-3b/RIP3-KD, and DA1-3b/RIP3-RHIM cells, which were analyzed 10 h after the addition of 1 mM IPTG. Representative images of live DA1-3b/GFP and DA1-3b/RIP3-RHIM cells and apoptotic DA1-3b/RIP3-WT and DA1-3b/RIP3-KD cells are shown. The images are shown at × 7000 magnification
