Anti mettl3 15073 1 ap

The antibodies used in this study were: anti-GAPDH (clone 4G5, AbD serotec, Atlanta, GA), anti-FLAG (F-3165, Sigma-Aldrich), anti-FTO (ab124892, Abcam, Cambridge, MA), anti- AlkBH5 (HPA007196, Sigma-Aldrich), anti-METTL3 (15073-1-AP, Proteintech Group, Rosemont, IL), anti-METTL14 (HPA038002, Sigma-Aldrich), anti-YTHDF1 (ab99080; Abcam), anti-YTHDF2 (ABE542, EMD Millipore, Billerica, MA), anti-YTHDF3 (ab103328; Abcam), and anti-HIV-1 Gag (clone #24–2, the NIH AIDS Reagent Program). Cells were harvested and lysed in cell lysis buffer (Cell Signalling, Beverly, MA) supplemented with protease inhibitor cocktails (Sigma-Aldrich). Immunoblotting was performed as described (St Gelais et al., 2015 (link)). Detection of GAPDH (glyceraldehyde-3-phosphate dehydrogenase) expression was used as a loading control.

The primary antibodies of anti-hnRNPL (ab6106), IgG (ab171870) were from Abcam (Cambridge, MA, USA); the primary antibodies of anti-ALKBH5 (16837-1-AP), anti-β-actin (66009-1-g) and anti-METTL3 (15073-1-AP) were from Proteintech (Wuhan, China); the primary antibodies of anti-Bcl2 (3498S), anti-caspase3 (9665S), anti-caspase9 (9502S) and anti-Ubiquitin (3933S) were from Cell Signaling Technology (Danvers, MA, USA). Cisplatin, Actinomycin D, Cycloheximide and MG132 were purchased from MedChemExpress (Monmouth Junction, NJ, USA). Antisense LNA^TM GapmeRs for CASC8 were synthesized by QIAGEN (Frankfurt, Germany). siRNAs for CASC8, hnRNPL, and ALKBH5 were synthesized by Ribobio (Guangzhou, China).

Anti-FBXO31 antibody (#86137; 1:1000) antibody was purchased from Abcam. Anti-AuroraA antibody (#14475; 1:1,000), anti-Slug antibody (#9585; 1:1000), anti-C-myc antibody (#5605; 1:1000), anti-p-p44/42 MAPK (Erk1/2) (#4370; 1:1000) and anti-tubulin antibody (2128 S; 1:2000) antibodies were all obtained from Cell Signaling Technology. Anti-SIRT2 (66410-1-Ig; 1:1500), anti-METTL3 (15073-1-AP; 1:1500), anti-METTL14 (26158-1-Ap; 1:1500), Anti-Flag tag (20543-1-AP; 1:1500), anti-HA tag (51064-2-AP; 1:1000), Anti-β-actin (20536-1-AP; 1:2000) and anti-His tag (66005-1-Ig; 1:1,000) antibodies were all bought from Proteintech. Peroxidase-conjugated anti-mouse secondary antibody (70-GAM007, 1:5000) and peroxidase-conjugated anti-rabbit secondary antibody (70-GAR0072; 1:5,000) were purchased from MultiSciences Company.