Fig. 5

OLMALINC, an upregulated lncRNA in osteogenic medium induced human renal interstitial fibroblasts (hRIFs), directly bound to OCT4 and elevated OCT4 protein level. A OCT4-RIP-Seq was performed in hRIFs induced with osteogenic medium for a week. Data were expressed as mean adjusted TPM (log2); n = 3. Red dots showed the transcripts of lncRNAs with enrichment in OCT4-RIP more than fourfold to input. B RNA sequencing for total RNA was performed to identify differentially expressed lncRNAs between osteogenic group (7 days; n = 3) and normal group (7 days; n = 3). Red dots showed the transcripts of lncRNAs with fold change > 2 and Q value < 0.05. C Venn diagram illustrated the intersection of OCT4 binding lncRNAs identified by RIP-Seq and upregulated lncRNAs identified by RNA profiling. D–E RIP-qPCR was performed to verify those OCT4 binding lnRNAs with top 5 enrichment fold among the intersection of OCT4-RIP-Seq and RNA sequencing for total RNA; n = 3. F QRT-PCR analysis of the expression of TARID, OLMALINC, and LOC100130872 in hRIFs cultured in either osteogenic medium (n = 3) or normal medium(n = 3) for 7 days. G, H Agarose gel electrophoresis showed the PCR products of 5′ and 3′ RACE for OLMALINC, and the sequences of 5′ and 3′ tail end were shown. I The location of OLMALINC determined by fluorescence in situ hybridization (FISH); n = 3. J QRT-PCR analysis of OLMALINC in hRIFs with overexpressed or silenced OCT4; n = 3. K, L QRT-PCR and immunoblotting (IB) analysis of OCT4 in hRIFs with overexpressed or silenced OLMALINC; n = 3