Figure 4.

Circ-PRKCI regulated PLCB1 by sequestering miR-1324.

(a)Putative miR-1324 binding sites in the 3′-UTR of PLCB1 mRNA was showed. The sequence in the PLCB1 3′-UTR at the complementary sites of miR-1324 was mutated. (b) Dual luciferase reporter showed significant decrease of luciferase activity of the PLCB1 wild-type and luciferase activity was restored by the PLCB1 mutant sequence. (c) The expression of PLCB1 in HSCR tissues and control tissues. PLCB1 was distinctly reduced in patient tissues compared with control tissues. (d) Western blot analysis showed that the protein level of PLCB1 was down-regulated in 3 pairs of HSCR tissues compared to normal control samples. (e) Bivariate correlation analysis of the relationship between circ-PRKCI and PLCB1 expression level. (f) Circ-PRKCI was inversely correlated with miR-1324 expression in the same paired intestinal tissues (P = 0.0021, Spearman). (g) PLCB1 was negatively correlated with the expression level of miR-1324 in the same paired intestinal tissues (P = 0.0089, Spearman).