Fig. 3

The NMRK2 protein expression was suppressed by NONO fragment of NONO-TFE3 fusion in NONO-TFE3 rRCC. A UOK109 cells were transfected with lentivirus shRNA (NC) or shRNA (TFE3). The expression of NMRK2 protein was measured by Western Blotting. β-actin was used as the internal reference. B HEK293T cells were transfected with vector plasmid or TFE3 (6–10 exon)-Flag plasmid, and the NMRK2 protein expression was measured by Western Blotting. β-actin was used as the internal reference. C HEK293T cells were transfected with vector, NONO (1–9 exon)-Flag, TFE3 (6–10 exon)-Flag, or NONO-TFE3-Flag plasmid, and the NMRK2 protein expression was measured by Western Blotting. β-actin was used as the internal reference. D UOK109 cells were transfected with lentivirus shRNA (NC) or shRNA (TFE3). The expression of NMRK2 NM or NMRK2 NR was measured by qPCR. 18S rRNA was used as the reference gene. E The subcellular distribution of NMRK2 mRNA was detected with the FISH assay. The U6 probe and 18S rRNA probe were used for positive control for the nucleus and cytoplasm locations, respectively. F The enrichment of m7G in NMRK2 mRNA was assessed by RIP assay. Rabbit IgG was used as a negative control. G A schematic representation of dCas13b-HA RIP assay. H The Wayne diagram showed the intersection of the RIP-seq dataset and the NONO-TFE3 fusion protein ChIP-seq dataset. I The volcano map showed the overlapped genes between the RIP-seq dataset and the NONO-TFE3 fusion protein ChIP-seq dataset. J UOK109 cells were transfected with Lentivirus shRNA (NC), shRNA (MALAT1)-1, shRNA (MALAT1)-2, or shRNA (MALAT1)-3. The NMRK2 protein expression was detected by Western Blotting. β-actin was used as the internal reference. K And the results were quantified with Image J. L The ribosomes in UOK109 cells from each group were extracted and purified with a Ribosome Extraction Kit. The total RNA in ribosomes was isolated with RNA Easy Isolation Reagent, and the expression of NMRK2 mRNA was measured by qPCR. 18S rRNA was used as the reference gene. M A schematic representation of the translation repression machinery by lnc-MALAT1. Data are presented as the mean ± SEM. *P < 0.05, ***P < 0.001