Sh ythdf1 1

HEK-293 T cells were used to produce lentivirus particles. Briefly, pLKO.1 puro-sh-YTHDF1#1/sh-YTHDF1#2 or pLKO.1 puro control vector, helper vector pxPAX2 and envelope vector pMD2.G were transfected into HEK-293 T cells using Lipofectamine 2000 (Thermo Fisher Scientific, US). Supernatants containing lentivirus particles were collected and filtered after 48 h transfection. 1 * 10⁴ cells were incubated with 1 ml supernatants for 24 h and were selected using 4 μg/ml puromycin. 1 μg/μl puromycin was used as a selective pressure to get stably transfected cell lines. sh-YTHDF1#1 and sh-YTHDF1#2 were synthesized by GenePharma (CN). Oligo sequence of sh-YTHDF1#1: 5′-CGGTGGGACAAATGTGAACAT-3′; sh-YTHDF1#2: 5′-CCCGAAAGAGTTTGAGTGGAA-3′; sh-YTHDF1-3: 5′-GTTCGTTACATCAGAAGGATA-3′. sh-FOXM1#1: 5′-GCTGGGATCAAGATTATTA-3′; sh-FOXM1#2: 5′-GGCTGCACTATCAACAATA-3′. Scramble shRNA was purchased from Addgene (#1864, US).

All cell transfections were performed by Lipofectamine 2000 reagent (Invitrogen, USA), including Sh-NC, Sh-METTL3#1, Sh-METTL3#2; Vector-OE, METTL3-OE; lncRNA METTL4-2-OE; Sh-NC, Sh-YTHDF1#1; and Sh-YTHDF1#2; (GenePharma, Shanghai, China). Cells were harvested at 48 h post-transfection for the following experiments.