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. 2021 Sep 23;22(19):10247. doi: 10.3390/ijms221910247

Figure 5.

Figure 5

I3C inhibits NEDD4-1 expression and represses temozolomide-resistant GBM cell line Nrf2-induced antioxidant response. (A) Protein expression of NEDD4-1, PTEN, p-AKT, AKT, NRF2, and HO-1 was evaluated after inducing oxidative stress with TBHp and treatment with I3C. (B) Effect of various doses of I3C on antioxidant enzyme activity in U87MGR cells. (C) Flow cytometry analysis of TBHp-induced ROS formation in U87MGR cells with/without I3C treatment using the ROS-sensitive fluorometric probe DCFDA. Relative ROS production in all cells tested normalized to TBHp + vehicle group. (D) Cell apoptosis rate assay using annexin V/PI staining of U87MGR cells treated with I3C and/or temozolomide. (E) Cell viability assay of U87MGR cells treated with I3C and/or temozolomide. Graph bars are the mean ± SEM of three independent experiments. * p < 0.05, ** p < 0.01, *** p < 0.001.