Fig. 3: tRF-Ser directly binds to CNBP and restricts its nuclear localization.

A RNA pull-down combined with silver staining to identify differentially bound proteins with tRF-Ser in MKN-45 cells. B List of the top six candidate binding proteins identified from the differential band, ranked by score. C Validation of the tRF-Ser-CNBP/RPS16/CST6 interaction by RNA pull-down assays in MKN-45 and HGC-27 cells. D The unique peptide of CNBP identified by mass spectrometry assay. E Validation of the tRF-Ser-CNBP interaction by RIP assays in MKN-45 and HGC-27 cells. F Validation of the tRF-Ser-CNBP interaction by CLIP assays in MKN-45 and HGC-27 cells. G–I WB (G, H) and IF (I) assays showed tRF-Ser overexpression inhibited CNBP nuclear accumulation in MKN-45 cells. J–L WB (J, K) and IF (L) assays showed tRF-Ser knockdown promoted CNBP nuclear accumulation in MKN-45 cells. M–P CNBP knockdown reversed the pro-tumorigenic effects of tRF-Ser knockdown. Functional assays including CCK-8 (M), colony formation (N), Transwell invasion (O), and cell cycle analysis by flow cytometry (P) in GC cells. Data are expressed as mean ± SD. (Student′s t-test, **p < 0.01, and ***p < 0.001). ns means no significant.