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. 2019 Jul 29;20(15):3700. doi: 10.3390/ijms20153700

Figure 2.

Figure 2

CHK1 inhibition activates the ataxia telangiectasia mutated serine/threonine kinase (ATM)-p53-p21 axis of the DNA damage response (DDR) pathway. (A) Quantitative RT-PCR analysis of p53, MDM2, and p21 mRNA levels in NB-39-nu cells transfected with 10 nM p53-specific or control siRNA for 24 h and then treated with 1 μM CHK1i (PF-47736) or DMSO for 24 h. Data are presented as the mean ± SD of triplicates and are normalized to ACTB levels. * p < 0.05. (B) Immunoblot analysis of the indicated proteins in NB-39-nu cells treated for 24 h with 1 μM CHK1i or DMSO (NT). β-actin was used as a loading control. (C) Indirect immunofluorescence of NB-39-nu cells treated for 24 h with 1 μM PF-477736 or DMSO. Cells were stained with polyclonal anti-p-p53-Ser15 (red) and 4′,6-diamidino-2-phenylindole (DAPI, nuclear DNA, blue).