Fig. 5: High glucose-enhanced cell death reverts to caspase-dependent death in the absence of RIP1.

a RIP1 was mutated in U937 cells using CRISPR–Cas9. Mutant cells (Mut) do not express RIP1 protein compared to wild-type (WT) and cells exposed to a non-targeting CRISPR control (NTC). b Mutation of RIP1 does not prevent the enhancement of TNF/CHX-induced cell death in high glucose conditions. c High glucose-enhanced death of RIP1 mutant cells treated with TNF-α/CHX is not prevented by the RIP1 inhibitor, nec-1s. High glucose-enhanced death of RIP1 mutant cells treated with TNF-α/CHX is prevented by d the pan-caspase inhibitor, zVAD, e the caspase-8 inhibitor, zIETD, and f the caspase-3 inhibitor, zDEVD. g Flow cytometry of RIP mutant cells and non-targeting control (NTC) following activation of TNF/CHX-induced apoptosis. Annexin V staining (apoptosis marker) of NTC cells decreases in high glucose following TNF/CHX treatment. Annexin V staining of RIP1 mutant cells does not decrease in high glucose. Two-way ANOVA, *p < 0.05, ***p < 0.001