Fig. 2.

PCBP1 destabilizes BECN1 mRNA.

(A) BENC1 mRNA 3′-UTR sequence and putative PCBP1 binding region were analyzed using RBP map and UCSC genome browser. (B–C) The fragments of BECN1 3′-UTR mRNA were produced by deletion and site-directed mutation PCR and inserted into the pGL4.54 vector. (D) Luciferase activity was measured in the HEK293FT cell line transfected with BECN1 fragment inserted vector and pcDNA3.1-PCBP1 overexpression vector. (E) mRNA affinity isolation assay was performed with the biotinylated transcripts of the BECN1 mRNA 3′-UTR, and then PCBP1 binding to the different fragments of the BECN1 mRNA 3′-UTR was determined by immunoblotting. The error bars represent standard errors from three independent experiments. ns; not significant, *P < 0.05, **P < 0.01, ***P < 0.001 between vector and PCBP1 plasmid.