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. 2019 Jan 30;9(4):1047–1065. doi: 10.7150/thno.29820

Figure 6.

Figure 6

Mitochondrial apoptosis signaling pathways. (A) Mitochondrial membrane potential (Δψm) in cells with the treatment of NP-cRGD+US, cisplatin, Pt(IV), Pt(IV) NPs, Pt(IV) NPs+US, Pt(IV) NP-cRGD and Pt(IV) NP-cRGD+US (30 μM eq.) for 48 h, as detected by the ratio of fluorescent intensity ([green]/[red]) of JC-1. (B) Immunohistochemically stained images of cytochrome c translocated from the mitochondria to the cytosol. Original magnification, 200×; Scale bar, 100 μm. (C, D) Activities of caspase 9 and caspase 3 as determined by a microplate reader. (E) Western blotting analyses of Bcl-2, Bax, Cleaved caspase 9 and Cytochrome c after different drug formulations incubations. (F, G) Cell apoptosis of the SKOV3 cells treated with NP-cRGD+US, cisplatin, Pt(IV), Pt(IV) NPs, Pt(IV) NPs+US, Pt(IV) NP-cRGD and Pt(IV) NP-cRGD+US (30 μM eq.) for 48 h, using the Annexin V and PI kit to detect the apoptosis ratio through the flow cytometry analysis, along with the statistics of cell apoptosis ratio, EA: early apoptosis; LA: late apoptosis; TA: total apoptosis. The data are presented as the mean ± SD of three independent experiments. Statistical significance in (A), (C) and (D) was calculated by one-way ANOVA with Tukey's post hoc test. Statistical significance in (G) was calculated by two-way ANOVA with Tukey's post hoc test. *P < 0.05, **P < 0.01, ***P < 0.005, NS indicates P >0.05.