Fig 2: SeRNA-NPCM interacts with hnRNPR and ACTA1.A Western blotting analysis on the association of seRNA-NPCM with hnRNPR and ACTA1 in S18 cells. GAPDH served negative control. B RNA immunoprecipitation (RIP) with antibody against ACTA1 or hnRNPR in non-crosslinked S18 cells transfected with seRNA-NPCM and qRT–PCR analysis of retrieved RNAs. IgG-bound RNA served as a negative control, U1 as a positive control. C Western blotting assay confirming hnRNPR and ACTA1 enrich target protein in RIP. IgG was a negative control. D The indicated deletion fragments of seRNA-NPCM and its interacting region with ACTA1 or hnRNPR. AS stands for antisense. E, F S18 cells were transfected with shRNAs targeting ACTA1, hnRNPR or scrambled negative control. The expression of target genes was measured by qRT–PCR (n = 4 independent experiments). Data are expressed as mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001.

Fig 3: SeRNA-NPCM harbors oncogenic properties through hnRNPR-mediated NDRG1 expression.A NDRG1 expression in the cells transfected with vector, seRNA-NPCM, scramble shRNA or sh-seRNA-NPCM, and those co-transfected with shNDRG1 or NDRG1 (top). NDRG1 transcription in the cells transfected with vector, seRNA-NPCM, shscramble or sh-seRNA-NPCM, and those co-transfected with shNDRG1 or NDRG1 (bottom). B–D Migration and invasion capability of the cells transfected with vector, seRNA-NPCM, shscramble or sh-seRNA-NPCM, and those co-transfected with shNDRG1 or NDRG1 (n = 4 independent experiments). E hnRNPR expression in the cells transfected with vector, seRNA-NPCM, scramble shRNA or sh-seRNA-NPCM, and those co-transfected with sh-hnRNPR or hnRNPR. F NDRG1 transcription in S26 cells transfected with vector, seRNA-NPCM, shscramble or sh-seRNA-NPCM, and those co-transfected with sh-hnRNPR or hnRNPR. Data are expressed as mean ± SEM; *P < 0.05, **P < 0.01, ***P < 0.001.