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. 2018 May 1;9(33):23029–23046. doi: 10.18632/oncotarget.25196

Figure 6. p53 plays a major role in killing normal skin cells following exposure to pladienolide B.

Figure 6

(A) 24 hours prior to the addition of pladienolide B (PD) NHF and RDEBK were transfected with three individual siRNAs targeting all known isoforms of p53. Cell death was analysed by real-time imaging 72 hours after pladienolide B addition. Knockdown of p53 reduced sensitivity to pladienolide B. (B) NHF were transfected with three individual p53 siRNAs. 24 hours after transfection cells were treated with carrier (-) or 100 nM pladienolide B (+). Cells were harvested at the time of drug addition (T0) and 24 hours later. p53 was efficiently knocked down by all of the siRNAs. (C) NHF, RDEBK and NHK were treated with pladienolide B (PD) for 24 hours. Protein expression was analysed by western blotting. Strong p53 upregulation was associated with reduced full-length MDM2 (MDM2FL) protein expression. (D) NHF and NHK cells were treated with pladienolide B (PD) for 24 hours. PCR was carried out with primers complementary to the indicated exons (E). 10 and 100 nM pladienolide B altered the ratio of alternatively spliced forms of MDMX mRNA and 100 nM pladienolide B dramatically interfered with MDM2 mRNA splicing.