Research Paper Volume 13, Issue 2 pp 2822—2850

Figure 1. Silencing Rac1, p22-phox, p47-phox, or p67-phox reduces OGD-induced ROS production. (A) Intracellular ROS production, as measured by flow cytometry analysis of DCFH-DA probe fluorescence. (B) ROS production after 0 h, 12 h and 18 h OGD treatment. ROS production was notably increased after an OGD. (N.S. no significance, ***P < 0.001, statistically analyzed by one-way ANOVA followed by the Bonferroni correction, n = 3). (C–G) Rac1, p22-phox, p47-phox, and p67-phox protein expressions in wild type BMMSCs after exposing to different durations of OGD (0 h, 12 h, or 18 h). (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, statistically analyzed by one-way ANOVA followed by the Bonferroni correction, n = 4). (H–K) Representation of flow cytometric histograms of different transfected BMMSCs and parental BMMSCs with or without an OGD 12 h stimulation. (L) Statistical graph showing ROS production of different BMMSCs cell lines under different hypoxia stimulation conditions. In comparison to shLuci, the other lentiviruses showed reduced ROS production to varying degrees. (N.S. no significance versus BMMSCs group, #P < 0.05 versus BMMSCs group, **P < 0.01 versus shLuci+OGD group, ***P < 0.001 versus shLuci+OGD group, statistically analyzed by one-way ANOVA followed by the Bonferroni correction, n = 3). Data are presented as mean ± SD.