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. 2020 Dec 8;2020:3042636. doi: 10.1155/2020/3042636

Figure 5.

Figure 5

Effects of 10-HDA on cell cycle distribution and cell cycle checkpoint-related proteins of human lung cancer cells. (a) The cells were treated with 10-HDA for different time periods (3, 6, 12, and 24 h). A549 cells were stained with 100 μL RNase A and 400 μL PI; DNA content was analyzed for cell cycle phase distribution by flow cytometry. (b) Western blotting with antibodies against p-AKT, AKT, p21, p27, CDK2/4/6, and cyclin D1/E was normalized to α-tubulin. (c) A549 cells were cultured with 30 μM 10-HDA or 0.25 μM (20 μL/mL) NAC for 24 h and detected by flow cytometric analysis. (d) A549 cells were pretreated with 0.25 μM (20 μL/mL) and NAC for 30 min, followed by treatment with 10-HDA for 24 h. The expression levels of p-AKT, AKT, p21, p27, CDK2/4/6, and cyclin D1/E were normalized to α-tubulin. The expression levels of proteins were analyzed by ImageJ software. ∗p < 0.05, ∗∗p < 0.01, and ∗∗∗p < 0.001 vs. the control group and the NAC+10-HDA group.