Fig. 4

PEBP1P2 inhibits the decay of PEBP1 mRNA via RNA modification. a Venn diagram of the identified gene candidates which could interact with PEBP1 mRNA and PEBP1P2 predicted by ENCORI database. b The mRNA level of PEBP1 after transfection with indicated shRNAs was detected by real-time PCR, respectively. c Luciferase activity of PEBP1-full-length was measured after transfected with indicated lentivirus. d The RNA pulldown assay, MS2-RIP, and western blot were performed to determine whether protein candidates could bind PEBP1 RNA. e PEBP1 mRNA enrichment in the complex of protein candidates was assessed by RIP assay and real-time PCR, and the mRNA enrichment was quantified by IgG control. f–h Luciferase activity of PEBP1-full-length and the PEBP1-mutation with the depletion of indicated regions was measured after being treated with indicated lentivirus. i The ELAVL1 binding capacity to PEBP1 mRNA with indicated mutations was validated by MS2-RIP, RNA pulldown assay, and western blot. j Real-time PCR was used to determine the stability of PEBP1 and PEBP1 with indicated mutations and GAPDH transcripts in 786-O cells transfected with PEBP1P2-shRNA. k, l Following transfection with indicated lentiviruses, PEBP1 mRNA levels were determined by real-time PCR. m Real-time PCR was used to assess the stability of PEBP1 mRNA in 786-O cells transfected with indicated lentiviruses. n ELAVL1 and YBX1 were tested for interaction using the CoIP assay. o The interaction between YBX1 and PEBP1 mRNA was elevated using MS2-RIP, RNA pulldown assay, and western blot. p The mRNA levels of YBX1 and PEBP1 after transfection with shYBX1 were measured by real-time PCR. q–s After transfection with indicated lentiviruses, MS2-RIP, RNA pulldown assay, and western blot was performed to determine whether YBX1 and PEBP1 transcripts interact. The data are presented as the mean ± SD, ***P < 0.001, n.s. no significance