Fig. 2

NAT10 depletion inhibits GC cell proliferation and in vivo tumor growth. A Schematic of NAT10 with its known domains. Arrows indicate point mutations. B Western blot analysis was performed to confirm the level of NAT10 in control AGS cells, NAT10-knockout cells, knockout cells rescued with stable expression of wild-type or mutant NAT10, and BGC823 cells stably expressing NAT10 shRNAs or control shRNA. C-F The ac4C mRNA levels were tested by HPLC–MS/MS (C) and ac4C dot blot (D) analyses, and proliferation capacities were detected by CCK-8 (E) and colony formation (F) assays in the above cells. G The cell cycle distribution was assessed in the indicated cells by flow cytometry. H and I NAT10 knockout inhibited subcutaneous tumor growth (H) and the formation of tumor nodules in the peritoneal cavity (I), while overexpression of wild-type NAT10, but not of the K290A or G641E mutants, offset these effects (n = 5 mice/group). CTR, control; KO, NAT10 knockout; shCTR, control shRNA; shNAT10, NAT10 shRNA. Error bars indicate the SD. *P < 0.05, **P < 0.01, *** P < 0.001 (two-tailed t-test)