Figure 2.
miR-339-5p participates in regulating p66Shc expression. (A) Hierarchical clustering analysis of 74 miRNAs that were significantly downregulated in intestinal I/R samples compared with sham samples (greater than 2.0-fold change, P< 0.05). The expression levels are represented with shades of red and green, which indicate expression above and below the median expression level across all samples (log2 scale, from -5.0 to +5.0), respectively. (B) A conserved binding site for miR-339-5p exists in the 3'UTR of p66Shc. (C) The expression levels of miR-339-5p in mouse I/R intestines (**P<0.01 compared with the sham group) (left) and H/R-treated Caco-2 cells (**P<0.01 compared with the control group) (right) were analyzed by qRT-PCR (n=6). (D-E) Caco-2 cells were transfected with ago-339, ant-339 or negative controls. (D) p66Shc protein expression (n=3). (E) p66Shc mRNA expression (n=6). (F) The miR-339-5p binding site was mutated in the p66Shc 3'UTR. BS, binding site. (G) Luciferase activity of luc-p66Shc-wt or luc-p66Shc-mut in Caco-2 cells cotransfected with ago-339 or ago-NC (n=6). **P<0.01. The values represent the mean ± SD.