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. 2020 Oct 13;11(10):852. doi: 10.1038/s41419-020-03082-9

Fig. 4. RPS27L knockdown impairs ICL repair and triggers MMC sensitization.

Fig. 4

A, B RPS27L knockdown reduces ICL repair upon MMC treatment. A549 cells, infected with indicated lentivirus, were treated with MMC at 1 µM for indicated periods of time, followed by immunofluorescent staining with FANCD2 (A) or γH2AX (B) Ab (top). Scale bar = 20 μm. The cells with foci in five random fields were counted (bottom). Shown are mean ± SEM from three independent experiments; *p < 0.05; **p < 0.01. C RPS27L knockdown impairs ICL repair. H1299 cells, infected with indicated lentivirus, were transfected with plasmid reporter substrates with a site-specific ICL for 24 h, and then subjected to luciferase reporter reactivation assay. Shown are mean ± SEM from three independent experiments; **p < 0.01. D, E RPS27L knockdown sensitizes lung cancer cells to MMC treatment. A549 cells were infected with indicated lentivirus for 48 h, and then treated with various concentrations of MMC for 48 h, followed by MTT assay (D). H1299 cells were infected with indicated lentivirus, and then treated with MMC for 24 h, followed by clonogenic survival assay (E). Shown are mean ± SEM from three independent experiments. F CQ treatment partially abrogates MMC sensitization upon RPS27L knockdown. A549 cells were transfected indicated siRNA oligos for 48 h, and then treated with various concentrations of MMC in the absence or presence of CQ (25 µM) for 24 h, followed by CCK-8 assay. Shown are mean ± SEM from three independent experiments; *p < 0.05, **p < 0.01, ***p < 0.001, ns, no significance.