Scrambled rna scrna

All cells were transfected using jetPRIME reagent (Cat. 114-15, Polyplus Transfection, Inc., NY, USA) at a ratio of DNA to reagent of 1:3 according to the manufacturer’s protocol. In our previous study, the pcDNA6-V5-His-tag expression vector was purchased from Invitrogen (Cat. V22020, Carlsbad, CA) and pFOXM1 was provided by Dr. Ju-Seog Lee (MD Anderson Cancer Centre, TX, USA) [32 (link)]. Scrambled RNA (scRNA) was purchased from GenePharma (Shanghai Co., Ltd., China), and siFOXM1 was synthesized at the ST Pharm Oligo center (ST Pharm, Seoul, Korea).

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.