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. 2022 Jan 14;8:26. doi: 10.1038/s41420-021-00791-8

Fig. 2. Radiation-induced apoptosis and DNA damage of MLE-12 were reduced by knockdown and inhibition of Rac1.

Fig. 2

A CCK-8 drug toxicity test of NSC23766 for MLE-12. Cells were treated with PBS or different doses of NSC23766 (1, 10, 100, and 500 μM) for 24 h and then tested with CCK-8. B, C MLE-12 cells were pretreated with PBS or different doses of NSC23766 (10, 50, and 100 μM) for 2 h before 0 Gy (no radiation) or 10 Gy of radiation. The apoptosis ratio was detected by the Annexin V/PI method with a flow cytometry 24 h after radiation. D, E Cells were pretreated with PBS or 100 μM NSC23766 for 2 h and collected at designed time points after 10 Gy of radiation for WB detection of the expression of Caspase-3 and Bax. Rac1 knockdown (Rac1-SH) and control (Rac1-shNC) MLE-12 cell lines were constructed using Lentivirus vector. Cells were collected before (0 Gy) and at designed time points after 10 Gy of radiation. F, G Apoptosis ratio was detected using Annexin V/PI method by a flow cytometer. H, I Apoptosis-related proteins, Bax and Caspase-3 were detected by WB analysis. J, K The expression of DNA damage-repair related proteins, p-ATR, p-Chk1, and γ-H2AX were also tested by WB analysis. *** represented P < 0.001 between the two groups. The error value was expressed as mean ± SEM. The experiment was repeated three times.