Fig. 4

Gly-tRF inhibits NDFIP2 3′ UTR luciferase reporter activity. A qRT-PCR was used to detect the mRNA expression of target genes predicted by miRDB in HCCLM3 cells transfected with Gly-tRF mimic. B Expression of NDFIP2 gene in 371 HCC tissues and 50 normal liver tissues from the TCGA database. C The expression of NDFIP2 is related to the prognosis of HCC Kaplan–Meier Plotter database. D The expression of NDFIP2 in HCC was verified by immunohistochemistry. E The sequence of Gly-tRF and the binding site of NDFIP2 3′UTR are shown. The sequence of the NDFIP2 mutant plasmid is also listed. F Luciferase activity of cells transfected with pGL6 empty vector(vector), NDFIP2 3′ UTR wild-type(wt) and mutant plasmids(mut) co-transfected with Gly-tRF NC-mimic and Gly-tRF mimic in HEK-293T cells. Renilla luciferase activity (R value) was performed to correct and standardize firefly fluorescence activity (F value). Statistical graph data is expressed by F/R of three independent experiments. F Western blotting performed to detect the level of NDFIP2 protein in HCC cells stably transfected with Gly-tRF NC-mimic and Gly-tRF mimic. *P < 0.05, **P < 0.01, ****P < 0.0001