Fig. 5From: MLKL regulates Cx43 ubiquitinational degradation and mediates neuronal necroptosis in ipsilateral thalamus after focal cortical infarctionInteraction of MLKL Ser454 with Cx43 can trigger the opening of Cx43 hemichannels, leading to increased intracellular Ca2+, and cell necroptosis. A, B) Western blots to detect MLKL and pMLKL polymers in SH-SY5Y cells post TSZ treatment, Cx43, MLKL(WT), and MLKL(S454A) transfection. C (a-h) Dye uptake assay to detect Cx43 hemichannels in SH-SY5Y cells post TSZ treatment, Cx43, MLKL(WT), and MLKL(S454A) transfection. C (i-p) Fluo-4 AM to detect cytoplasmic Ca2+ concentration in SH-SY5Y cells post TSZ treatment, Cx43, MLKL(WT), and MLKL(S454A) transfection (scale: 50 μm). D) The histogram presents the quantitative analyses of EB fluorescence intensity (n = 6 in each group). E) The histogram presents the quantitative analyses of Fluo-4 AM fluorescence intensity (n = 6 in each group). MLKL, mixed lineage kinase domain-like protein; Cx43, connexin 43; WT, wild type; MT, mutant; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; TSZ, TNFα + Smac mimetic + ZVAD-FMK; EB, ethidium bromideBack to article page