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. 2019 Sep 18;10(10):688. doi: 10.1038/s41419-019-1927-0

Fig. 6. CELF6 regulates the stability of p21 transcript.

Fig. 6

a HCT116 wild-type cells or b HCT116 p53−/− cells overexpressing empty vector or His-CELF6 were treated with actinomycin (Act D) at 5 μg/mL for different time points, samples were collected for quantitative PCR and immunoblotting analysis to detect the expression levels of p21. c HCT116 cells overexpressing His-CELF6 were lysed, the cell lysates were immunoprecipitated using either anti-His antibody or control IgG. CELF6-associated p21 RNA was validated by qPCR using specific primers (***p < 0.001). ACTB was used as a negative control. d HCT116 cells were transfected with His-CELF6 and p21 expression vector containing 5′UTR + ORF, 3′UTR + ORF, or 5′UTR + 3′UTR + ORF (full length), the expression level of p21 was determined by immunoblotting. e Schematic diagrams or pGL3 reporters containing different combinations of p21 transcript. HCT116 cells were cotransfected with various pGL3 reporter, pRL-TK and His-CELF6, luciferase activity was determined using a dual-luciferase reporter kit