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. 2021 Jun 14;9:660005. doi: 10.3389/fcell.2021.660005

FIGURE 3.

FIGURE 3

Combination of sorafenib and TMZ triggered mitochondrial damage and nuclear AIF aggregation. (A) After treating U251 cells with a combination of sorafenib and TMZ for 24 h, the mitochondrial membrane potential was stained with JC-1 and assessed via flow cytometry analysis. The results showed that the combination of sorafenib and TMZ decreased the mitochondrial membrane potential of U251 cells. (B) Western blot assay of Bcl-2, Bax, and Cyt c protein expression in U251 cells treated with the combination of sorafenib and TMZ for 24 h. Western blot assay demonstrated that the combination of sorafenib and TMZ upregulated the expression of Bax and Cyt c and downregulated the level of Bcl-2. (C) Representative images of U251 cells with confocal microscopy showed that the combination of sorafenib and TMZ increased the expression of cleaved caspase 3 (120×) (bar, 10 μm). (D) Western blot assay of the expression of cytoplasmic and nuclear AIF protein in U251 cells for 24 h. The results showed that the combination of sorafenib and TMZ increased the nuclear aggregation of AIF.