Research Paper Volume 16, Issue 5 pp 4591—4608

Figure 5. GBA3 reduced the occurrence of necroptosis. (A) EdU staining (green fluorescence) on cell proliferation, nuclei were counterstained blue with DAPI (Scale bars, 50 μm). (B) Cell proliferation was detected by measuring the optical density at 450 nm, P-value was calculated by 2-tailed Student’s t test (ns, not significant). (C) Percentage of apoptotic cell was assessed via flow cytometry, P-value was calculated by 2-tailed Student’s t test (ns, not significant; **, P < 0.01). (D) Immunoblotting detection of MLKL and pMLKL (Ser358) in cells. Protein expression was normalized to actin levels and shown as relative values. P-value was calculated by 2-tailed Student’s t test (*, P < 0.05). (E) The activity of caspase 8 was assessed by measuring the optical density at 405 nm. Incremental activity of caspase 8 was expressed as a percentage of control, P-value was calculated by 2-tailed Student’s t test (*, P < 0.05; ***, P < 0.001). (F) Phosphorylation level of MLKL in livers detected by immunohistochemistry. (G) The infiltrated level of immune cells, P-value was calculated by Mann-Whitney U test (*, P < 0.05; **, P < 0.01).