Fig. 6

WTAP was involved in the cell cycle by alleviating the expression of p21 and p27. a and b Cell cycle distribution was analyzed by flow cytometry in Huh7 (a) and Hep3B (b) cells where WTAP were silenced, with summary bar charts showing the percentage of cells in each phase; c and d RT-qPCR was utilized to explore alterations of p21 and p27 when WTAP was knocked down in Huh7 (c) and Hep3B (d), respectively; e Cell cycle-related proteins including p21, p27, CDC25C, CDK1, cyclin-A2 and cyclin-B1 were measured by western blot in the indicated cells where WTAP was knocked down (Huh7 and Hep3B) or overexpressed (SMMC-7721 and HCCLM3); f and g Flow cytometry indicated that ETS1 knockdown could reverse the G2/M arrest in WTAP-silenced MHCC97H (f) or HCCLM3 (g) cell; h The knockdown efficiency of ETS1 was verified followed by the detection of p21 and p27 expression via western blotting; i The ChIP assay was conducted in Huh7 and HCCLM3 cells to determine whether ETS1 could bind to the promoter of p21 and p27 (IP/input was calculated); j and k A rescue assay was performed with or without knockdown of WTAP or ETS1 to validate the retrieved role of ETS1 in WTAP-mediated events. Expression of p21 and p27 were detected at the RNA (j) and protein (k) levels