Figure 5. NF-κB mediates the inhibitory effect of M₃-mAChR on miR-376b-5p expression.

GAPDH as an internal control normalized NF-κB band for western blot. U6 as an internal control normalized miR-376b-5p for quantitative real-time RT-PCR (qRT-PCR). (A) Chromosomal organization of the miRNA-376b gene locus on rat chromosome 6q32 indicating three NF-κB binding sites in the pre-miR-376b gene promoter. (B) Choline activated NF-κB in H9c2 cells. Top, representative Western blots of total NF-κB; Middle, representative Western blots of nuclear NF-κB; bottom, quantitation of nuclear NF-κB as mean ±SEM Note: n = 4; *P<0.05 versus Control. (C) LPS activated NF-κB in H9c2 cells. Top, representative Western blots of total NF-κB; Middle, representative Western blots of nuclear NF-κB; bottom, quantitation of nuclear NF-κB as mean ±SEM. Note: n = 6; *P<0.05 versus Control. (D) LPS inhibited miR-376b-5p expression in H9c2 cells. MiR-376b-5p expression was determined by qRT-PCR. Values were expressed as mean ± SEM; n = 5; *P<0.05 vs. Control.