Fig 2: Cd treatment induced the loss of mitochondrial transmembrane potential and the up-regulation of proapoptotic protein BAX(A) BEAS-2B cells were sham-exposed or dosed with 30 µM CdCl2 for 18 h. JC-1 assay was conducted as described in “Materials and methods” for the determination of mitochondrial membrane integrity. (B) BEAS-2B cells were sham-exposed or dosed with increasing concentrations of CdCl2 for 36 h; cells were lysed; and protein extracts were subjected to western blot analysis using antibodies against BAX. The same blot was stripped and reprobed with the monoclonal ß-actin antibody to monitor the loading difference. The results are representative of three independent experiments.

Fig 3: As2O3 promotes cell death in high ERK8-expressing lung cancersDose- and time-dependent cytotoxicity of As2O3 towards H1299 cells. H1299 cells were sham-exposed or exposed to 5-20 µM of As2O3 for 36 h or 5 µM As2O3 for 12-48 h. (A) cell morphology of H1299 cells under light microscope. (B) The corresponding western blot of (A), cells were lysed and protein extracts were subjected to western blot analyses using antibodies against Bax, Caspase-9, and Caspase-3. After development, the membranes were stripped and reprobed with ß-actin to monitor the loading difference. (C) Stable shMock and shERK8 H1299 cells were sham-exposed or exposed to 5 µM As2O3 for 48 h, the percentage of cell viability was determined by MTS assay. *A significant difference of P < 0.05. (D) The corresponding western blot of (C) for the determination of marker proteins involved in apoptosis. The data are representative of three independent experiments.

Fig 4: miR-106a-5p overexpression and Foxo1 knockdown inhibits the proliferation and promotes the apoptosis of VSMCs. (A) CCK-8 was used to detect cell viability in VSMCs overexpressing miR-106a-5p or Foxo1 (**p < 0.01 miR-106a-5p vs. mimics NC, *p < 0.05 pcDNA3.1/Foxo1 vs. pcDNA3.1). (B) Edu assay was used to detect cell viability by FCM. (C) FCM detected the apoptosis of VSMCs. (D) Western blot analysis detected the expression of Bax, Bcl2, PCNA, and Caspase 3.

Fig 5: Western blotting of brain tissues.Reactive oxygen species may activate the mitochondrial pathway leading to execution of apoptosis. Compared with the control group, no difference in the expression of cytochrome c, Bcl-2, or Bax was observed. Caspase-3 activation and PARP cleavage served as a proxy of execution of apoptosis pathway; also, no significant between-group differences are noted. Data are presented as the median and third quartile. A through C, Comparisons of expression level of cytochrome c, Bcl-2, and Bax in brain tissue by Western blotting. D and E, Comparisons of ratios between full-length and cleaved caspase 3 or PARP in brain tissue by Western blotting.