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. 2017 Jan 27;7:41166. doi: 10.1038/srep41166

Figure 6. Impact of SMIT1 overexpression on NOX2 activation and ROS production.

Figure 6

Adult rat cardiomyocytes were infected with adenoviruses (24 h, 200 MOI) expressing SMIT1 (Ad-SMIT1) or β-galactosidase (Ad-Ctl). (A) SMIT1 mRNA level measured by qRT-PCR (n = 3). Data were normalized to HPRT1 and expressed as relative expression vs Ad-Ctl. (B) SMIT1 protein expression in plasma membrane fractions obtained after cellular fractionation. Full-length blots are presented in Supplementary Figure 6. (C) Quantification of picomoles myo-[3 H]inositol uptake per min and mg of proteins (n = 4). (D) p47phox translocation close to cav3 (n = 6) and (E) ROS production (n = 7) in response to increased glucose concentration (5–10 and 21 mM of glucose). (F) Gp91dstat and scrambled peptide were added 15 min prior to glucose (5 or 10 mM glucose). ROS production was quantified 2 h after change in glucose concentration (n = 3). Data are means ± SEM. Statistical analysis was by (A–C) Student’s t-test or (D,E,F) two-way ANOVA. $Indicates values statistically different from LG, p ≤ 0.05. *Indicates values statistically different from (A–E) Ad-Ctl, and (F) scr, p ≤ 0.05.