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. 2022 Jan 20;12:1049. doi: 10.1038/s41598-022-05013-4

Figure 5.

Figure 5

p53/RBL2 regulates response to PTX and irradiation. Alisertib sensitizes cells to PTX/irradiation partially dependent on p53. (A,B) H1666 and cells were transfected with control siRNA, RBL2 siRNA (pooled and single), p53 siRNA (pooled and single), and LIN37 siRNA (pooled and single), then treated with the indicated doses of PTX (μM) for 48 h or irradiation. Cells were allowed to recover in drug-free media for 2–4 weeks. % average (three technical replicate from one experiment) formed colonies are presented with SD indicated. Statistical analyses are shown in Supplementary Table S13. Reduced gene expression in siRNA transfected cells is shown in (C). (D) H460 cells were treated with PTX (1 μM) or irradiation (4 Gy) in the absence or presence of alisertib (ALS, 0.1 μM) for 24 h. Lysates were immunoblotted for the indicated proteins (original immunoblot images are shown in Supplementary Fig. S5, representative images of at least two independent experiments). (E) and (F) H460 cells were treated with PTX (5 μM) or irradiation with or without ALS (50 nM) for 48 h (E). H460 cells were transfected with control siRNA or INCENP siRNA and then treated with PTX (5 μM) or irradiation (F). Cells were allowed to recover in drug-free media for 2 weeks. % average (three technical replicate from one experiment) formed colonies are presented with SD indicated. Statistical analyses are shown in Supplementary Table S14 and S15. (G) p53+/+ and p53−/− A549 cells treated with PTX (1 μM) or irradiation (4 Gy) with or without ALS (0.1 μM) for 24 h. Lysates were immunoblotted for the indicated proteins (original immunoblot images are shown in Supplementary Fig. S5, representative images of at least two independent experiments). (H) p53+/+ and p53−/− A549 cells treated with PTX (5 μM) or irradiation (4 Gy) with or without ALS (0.1 μM) for 48 h. % average (three technical replicate from one experiment) formed colonies are presented with SD indicated. Statistical analyses are shown in Supplementary Table S16.