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. 2014 Dec 19;5:5812. doi: 10.1038/ncomms6812

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(a,b) Western blot analysis of total p53 and phospho-p53 (Serine 15) on HCT116 cells transfected with ASOs for PR-lncRNA-1 (a) or PR-lncRNA-10 depletion (b) and treated as indicated. GAPDH is shown as a loading control. MW mark indicates 50 kDa. (c) p53 ChIP of p53 direct target genes regulated by PR-lncRNA-1 and PR-lncRNA-10. ChIP enrichment of p53 and control IgG of the indicated loci in HCT116 cells after the transfection with ASO pool for PR-lncRNA-1, PR-lncRNA-10 or ASO control for 36 h and treatment with 5-FU for 12 h. GAPDH promoter was included as a negative control. The mean±s.d. of three biological replicates, and the significant differences relative to the condition ASO ctrl +5-FU are shown. (d) Relative expression levels of the p53 direct target genes regulated by PR-lncRNA-1 and PR-lncRNA-10 in HCT116 cells treated like in c and determined by qRT–PCR. Values are the average of three biological replicates, and the significant differences relative to the condition ASO ctrl +5-FU are shown. All graphs (c,d) represent the mean (±s.d.) of at least three independent experiments. Significance was determined by two-tailed unpaired t-test. *P<0.05 and **P<0.01.