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. Author manuscript; available in PMC: 2021 Sep 1.
Published in final edited form as: Phytother Res. 2020 May 4;34(9):2366–2384. doi: 10.1002/ptr.6687

Figure 4: Nef induces oxidative stress in cervical cancer cells.

Figure 4:

(A) ROS generation was detected by adding 2’,7’-dichlorofluorescin diacetate (DCFDA) fluorescent dye. The fluorescent green color indicated the generation of ROS in Nef (10, 25 and 50μM) treated cells compared to the untreated cells. Conversely, the effect of Nef was abrogated in the presence of antioxidant N-acetyl cysteine (NAC) which resulted in decreased ROS production. (B) Nef induces apoptosis in cervical cancer cells by Annexin-V-FITC assay. HeLa (left panel) and SiHa (right Panel) cells were treated with various concentrations of Nef (10, 25 and 50μM) for 48 hrs. Cells were stained with Annexin V-FITC to detect apoptotic cells and PI for nuclear stain. Results showed that Nef induced apoptosis in both HeLa and SiHa cells. (C) Cell cycle analysis by Flow cytometry: HeLa and SiHa cells treated with Nef (10, 25 and 50 μM) was subjected to flow cytometry for cell cycle analysis. Data showed that Nef induced potent G0/G1phase cell cycle arrest in both HeLa and SiHa cells.