Sh e2f1

E2F1 full length was cloned into the overexpression plasmid (pcDNA-3.1, oe-E2F1). MiR-423-5p mimics and inhibitor, sh-E2F1, sh-HIPK2, and control sh-NC were synthesized from Genepharma (Shanghai, China). Lipofectamine 3000 (Invitrogen, Missouri, USA) was utilized as the reagent for cell transfection. In brief, cells were cultured to ~ 70% confluence, and then construct was added together with Lipofectamine 3000 at a ratio of 1:1. Cells were harvested for further analysis 48 h after transfection.

In this study, lipopolysaccharide (LPS, #L2880) and MIF antagonist (ISO-1, #475837) were bought from Sigma-Aldrich (Missouri, USA). Cycloheximide (CHX, #HY-12320) and 3-Deazaadenosine (DAA, #HY-W013332) were purchased from MedChemExpress (MCE, New Jersey, USA). VX765 (#inh-vx765i) was purchased from Invivogen (California, USA). The sh-IGF2BP1, sh-METTL3, sh-MIF, sh-E2F1, and sh-NC were obtained from Genepharma company (Shanghai, China). Lentiviruses expressing sh-IGF2BP1 and nonsense control (NC) were purchased from Tsingke (Nanjing, China). The E2F1 overexpression vector was constructed using p-ENTER, while the overexpression plasmids for MIF or METTL3 were constructed in pcDNA3.1.