miR-382 Contributes to Renal Tubulointerstitial Fibrosis by Downregulating HSPD1

Figure 3

Interaction between miR-382 and HSPD1. (a) Both TGF-β1 and pre-miR-382 could repress the protein expression of HSPD1 in HK2 cells. (b) Anti-miR-382 treatment could partially reverse the downregulation of HSPD1 that is induced by TGF-β1 in HK2 cells. (c) MiR-382 interacted with the 3′-UTR of HSPD1. HeLa cells at ~80% confluency were cotransfected with a 3′-UTR segment of HSPD1 (marked with luciferase reporter gene), a pRL-TK internal control plasmid, and control pre-miR oligonucleotides or the miR-382. Luciferase activity was significantly decreased by pre-miR-382, while in the mutated group, pre-miR-382 did not have significant effect on luciferase activity. compared with the control group; ^∇, ^# compared with the TGF-β1 group. ^▼ compared with the pre-NC+HSPD1 group; ^◇, ^◆ compared with the pre-382 + HSPD1 + anti-382 group. NC indicates negative control. Anti-382 indicates the group treated with anti-miR-382 oligos. Pre-382 indicates the group treated with pre-miR-382 oligos. Anti-NC indicates antinegative control. Pre-NC indicates prenegative control. UUO indicates unilateral ureteral obstruction. (a, b) Western blot; (c) 3′-UTR luciferase reporter assay and site-directed mutagenesis ().

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