Figure 1.

Evidence for sodium iodate-induced RPE necroptosis in vitro. (A) NaIO₃ (10 mM) treatment induces changes to cell membrane permeability in non-fixed ARPE-19 cells shown by staining with PI (a and b). Activation of RIPK3 after 2 h of 10 mM NaIO₃ treatment in RIPK3-GFP-transfected ARPE-19 cells (c and d). Mitochondrial network was tracked by ANT1-RFP and passive release of HMGB1 from the nucleus by HMGB1-YFP at 4 h after NaIO₃ treatment (e and f). (B) ARPE-19 cells were treated with 200 μM Nec-1, -5, -7; 3 μM GSK’872, or 25 μM resveratrol for 24 h before 10 mM NaIO₃ treatment. Cell viability was measured by MTT assay at 24 h. (C) ARPE-19 cells were treated with 50 μM z-VAD for 24 h before 10 mM NaIO₃ treatment. Cell viability was measured by MTT assay at 24 h later. (D) Activation of inflammasome was tracked by transfection of ASC-GFP-expressing plasmid in ARPE-19 cells. Alu RNA (5 ng) was used as a positive control for inflammasome activation (a and b). Treatment with 10 mM NaIO₃ did not induce inflammasome activation (c and d). (E) ARPE-19 cells were treated with 50 μM Ac-YVAD for 24 h before 10 mM NaIO₃ treatment. Cell viability was measured by MTT assay at 24 h later. *P<0.05; **P<0.01; ***P<0.001. The scale bar is 25 μM.