Figure 2. p66shc mediates CRIF1 deficiency-induced mitochondrial ROS and cytosolic ROS production in Crif1-silenced cells.

MS-1 cells were transfected with Crif1 siRNA for 48 h. (A) CRIF1 deficiency increased cytosolic ROS, measured using the Amplex Red H₂O₂ assay kit. Relative fluorescence of Amplex Red was used as a measure of the H₂O₂ released from the cells. (B) CRIF1 deficiency increased phosphorylation of p66shc (p-p66shc) on ser36. p-p66shc levels were measured by Western blotting, which are representative of three independent experiments, and quantified by densitometric analysis (B, lower panel). (C) and (D) p66shc mediated CRIF1 deficiency-induced ROS production. MS-1 cells were infected with Adp66shcRNAi or AdLacZ for 24 h, followed by transfection with Crif1 siRNA for 48 h. Data represent means ± SEM of three independent experiments, *P<0.05, ** P<0.01 compared with the control.