Sh foxm1

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).

pGL3-Basic and pRL-Renilla luciferase reporter plasmids were purchased from Promega (WI, USA). To generate the pGL3 Basic-FANCD2 promoter (from −3347 to –1), human genomic DNA was amplified by PCR using the indicated primer sets (Table S2). We purchased shRNA for FOXM1 from Sigma. The plasmids that stably expressed a shRNA against FOXM1 (shFOXM1) were established in a pLKO.1-TRC cloning vector (Addgene, MA, USA). We used a pLKO.1-puro nontarget shRNA plasmid (Sigma) for the negative control. pMD2.G and psPAX2 plasmids were purchased from Addgene (MA, USA). To construct pFANCD2 (FANCD2 overexpression vector), the coding sequence (CDS) of FANCD2 was inserted into the NheI/XhoI restriction enzyme sites of a pcDNA^TM6/V5-His A plasmid (Invitrogen, MA, USA). All constructs were verified using a DNA sequencing.
Scrambled RNA (scRNA) was purchased from Shanghai GenePharma (Shanghai, China). siFOXM1 (5’-GGACCACUUUCCCUACUUU-3’) was synthesized by the ST Pham Oligo Center (Korea). Plasmid and siRNA transfection were conducted using jetPRIME reagent (Polyplus, NY, USA) according to the manufacturer’s protocol.