Fig. 6
LncRNA like NMRK2 mRNA promoted the mitochondrial respiration of NONO-TFE3 rRCC by relieving the inhibitory effect of lnc-GAS5 on the TCA cycle. A UOK109 cells were co-transfected with dCas13b-HA and sg-Control/sg-NMRK2. The enrichment of lnc-GAS5 on NMRK2 under glucose deprivation for 24 h was assessed by RIP assay. Rabbit IgG was used as a negative control. B UOK109 cells were co-transfected with dCas13b-HA, sg-NMRK2, and mitochondria-RFP. The subcellular localization of lncRNA like NMRK2 mRNA was observed by the IF assay. Tom20 and β-tubulin were used as the positive and negative control for mitochondrial protein. C UOK109 cells were transfected with lentivirus shRNA (NC) or shRNA (NMRK2) and cultured with glucose deprivation for 24 h. The subcellular localization of lnc-GAS5 was observed by the FISH assay. D UOK109 cells were co-transfected with VN-MDH2 and VC-FH/VC-CS and cultured with glucose deprivation for 24 h. The binding between MDH2, FH, and CS was detected by the IF assay. E UOK109 cells were co-transfected with lentivirus shRNA (NC) or shRNA (NMRK2) and MDH2-Flag, and cultured in the setting of glucose deprivation for 24 h. The binding between MDH2, FH, and CS was detected by IP assay. F UOK109 cells were transfected with lentivirus shRNA (NC), shRNA (NMRK2), or shRNA (NMRK2 + GAS5) and cultured in the setting of glucose deprivation for 24 h. The mitochondrial respiration of cells in each group was measured by the Seahorse XF Extracellular Flux Analyzer with a Seahorse XF Cell Mito Stress Kit. G Schematic overview of the mechanism concerning lncRNA like NMRK2 mRNA and lnc-GAS5. Data are presented as the mean ± SEM. **P < 0.01