FIGURE 6.

MALAT1 sponged miR-320a-5p to promote osteogenic differentiation of human renal interstitial fibroblasts (hRIFs) via Runx2 upregulation. HRIFs were cotransfected with Len-sh-MALAT1 or Len-sh-ctrl and the miR-320a-5p inhibitor or NC-inhibitor; (A–C) qRT-PCR was performed to determine the relative mRNA expression levels of osteogenic markers (Runx2, OPN, OCN), and (D) the release of p-nitrophenol was measured to determine the relative activity of alkaline phosphatase (ALP) 7 days after osteogenic induction. HRIFs were cotransfected with Len-MALAT1 or Len-ctrl and the miR-320a-5p mimic or NC-mimic; (E–G) qRT-PCR was performed to determine the relative mRNA expression levels of osteogenic markers, and (H) the release of p-nitrophenol was measured to determine the relative activity of ALP 7 days after osteogenic induction. (I) Protein expression levels of osteogenic markers were determined by WB in cotransfected hRIFs 7 days after osteogenic induction. (J) qRT-PCR was used to determine the relative mRNA expression level of Runx2 in hRIFs transfected with Len-sh-Runx2 or Len-ctrl 7 days after osteogenic induction. (K) Alizarin Red staining in hRIFs transfected with Len-sh-Runx2 or Len-ctrl 14 days after osteogenic induction (100 ×). (L–N) Relative mRNA expression levels of OPN and OCN and relative activity of ALP in hRIFs transfected with Len-sh-Runx2 or Len-ctrl 7 days after osteogenic induction. (O) Protein expression levels of osteogenic markers were determined by WB in hRIFs transfected with Len-sh-Runx2 or Len-ctrl 7 days after osteogenic induction. GAPDH was used as the internal control. *P < 0.05; **P < 0.01; ***P < 0.001.