Fig. 6

OLMALINC elevated OCT4 protein level via inhibiting its ubiquitination. A, B Len-sh-ctrl or Len-sh-OLMALINC transfected human renal interstitial fibroblasts (hRIFs) were pretreated with CHX (10 μM), and OCT4 was determined by Immunoblotting (IB) at the indicated time points (n = 3). C, D HRIFs transfected with Len-sh-ctrl or Len-sh-OLMALINC were treated with the proteasome inhibitor MG-132 (10um/ml) for 8 h or not, and OCT4 was determined by IB (n = 3). E HRIFs transfected with Len-sh-ctrl or Len-sh-OLMALINC were treated with MG-132 for 8 h, and the cell lysis was collected for immunoprecipitation (IP; n = 4). The immune-precipitates of anti-OCT4 or IgG (negative control) were incubated with anti-ubiquitin. F IB analysis of OCN, BMP2 and RUNX2 in hRIFs with overexpressed or silenced OLMALINC 7 days after osteogenic induction (n = 3). G Alkaline phosphatase (ALP) activity of hRIFs with overexpressed or silenced OLMALINC 7 days after osteogenic induction (n = 3). H Alizarin Red Staining (ARS) for calcium nodes in hRIFs with overexpressed or silenced OLMALINC 14 days after osteogenic induction (n = 3). I Representative co-staining images of RNA FISH for OLMALINC (Red) and immunofluorescence for Vimentin (Green) in Randall’s Plaques (RP; n = 6) and normal renal papillae (NRP; n = 6). J QRT-PCR analysis of OLMALINC expression in RP (n = 28) and NRP (n = 28). K Liner regression analysis of OLMALINC expression and OCT4 protein density in RP (n = 18) and NRP (n = 18). L Linear regression analysis of OLMALINC expression and OCT4 mRNA expression in RP (n = 28) and NRP (n = 28)