Cyclohexamide

FG-4592 was obtained from either Selleckchem or MedChemExpress. Cyclohexamide was purchased from Abcam. siRNAs were obtained from Thermo Fisher Scientific, MG132 and 4-thiouridine, and N-ethylmaleimide were purchased from Sigma-Aldrich. Primary antibodies used in this study are ALKBH5 (Atlas Antibodies), β Actin (Sigma), HIF-1α (BD Bioscience), HIF-2α (NOVUS), HIF-1β (NOVUS), NDRG1 (CST), HBc (DAKO), anti-m⁶A polyclonal antibody (Synaptic Systems). HRP-conjugated secondary antibodies were purchased from DAKO.

Information on the sources, authentication and Mycoplasma testing of cell lines is provided in Supplementary Table 10. Cell lines were mycoplasma tested using the e-Myco mycoplasma PCR detection kit (iNtRON). HEK293A cell lines were used because they are an experimentally tractable and well-characterized model system46 (link). HEK293A cells were grown in DMEM (Gibco) with 10% FBS (Gibco). DLBCL lines were grown in RPMI (Gibco) with 10% FBS. All DLBCL cell lines were of the germinal centre B-cell subtype14 (link)47 (link). The MEF2B mutation status of the DLBCL cell lines was reported previously2 (link)14 (link) Transfected HEK293A were grown in 100 μg ml⁻¹ G418 (Invitrogen). Transduced DoHH2 were grown in 7.5 μg ml⁻¹ Blasticidin S (Invitrogen). Antibiotics were removed 24–48 h before harvesting cells. Cyclohexamide (Abcam) was used at 75 μg ml⁻¹. Cells for microarray and RNA-seq analysis were treated for 6 h with 1.07 μl ml⁻¹ of dimethyl sulfoxide (Fisher) immediately before RNA was collected, as a solvent-only control for drug treated cells (data from drug-treated cells was not presented).