Characterization of RNA Methylation Regulators in Adipogenesis

3T3-L1 pre-adipocytes were obtained from ATCC (Manassas, VA). The human FTO and mouse RUNX1T1 genes were cloned into pEGFP-C1B^{56 (link)}. Human SR plasmids were bought from OriGene Technologies (Rockville, MD). The expression constructs were generated using PCR and subcloned into pCS2+ vectors with an N-terminal FLAG tag^{57 (link)}. Polyclonal rabbit anti-FTO antibody was affinity-purified from rabbits immunized with 6×His-tagged full-length human FTO protein as previously reported^{9 (link)}. Polyclonal rabbit anti-ALKBH5 antibody was generated against synthesized peptide (RKYQEDSDERSD, 58-70 amino acids of human ALKBH5) by CWBio (Beijing) as previously reported^{34 (link)}. The primary antibodies were purchased from commercial sources: rabbit anti-METTL3 (MT-A70; 15073-1-AP, Proteintech Group); rabbit anti-RUNX1T1 (15494-1-AP, Proteintech); mouse anti-β-tubulin (t-5293, Sigma); rabbit anti-m⁶A (202003; Synaptic Systems); mouse anti-HA (cw0092B, CWbiotech); rabbit anti-Flag (F7425, Sigma); mouse anti-Flag (F1804, Sigma). The secondary antibodies used for immunoblotting and dot-blotting: anti-Rabbit IgG-HRP (p0448, dakocytomation); anti-Mouse IgG-HRP (p0161, dakocytomation).

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Zhao X., Yang Y., Sun B.F., Shi Y., Yang X., Xiao W., Hao Y.J., Ping X.L., Chen Y.S., Wang W.J., Jin K.X., Wang X., Huang C.M., Fu Y., Ge X.M., Song S.H., Jeong H.S., Yanagisawa H., Niu Y., Jia G.F., Wu W., Tong W.M., Okamoto A., He C., Danielsen J.M., Wang X.J, & Yang Y.G. (2014). FTO-dependent demethylation of N6-methyladenosine regulates mRNA splicing and is required for adipogenesis. Cell Research, 24(12), 1403-1419.

Publication 2014

3T3-L1 pre-adipocytes rabbit anti-m6A rabbit anti-Flag mouse anti-Flag anti-Rabbit IgG-HRP anti-Mouse IgG-HRP

3t3 l1 Adipocytes Amino acids Anti antibody Anti igg Antibodies Genes Human Human fto protein Mouse Mt a70 Peptide Plasmids Rabbit Rabbits Tubulin Vectors

Corresponding Organization :

Other organizations : Beijing Institute of Genomics, University of Chinese Academy of Sciences, Tsinghua University, The University of Tokyo, RIKEN Advanced Science Institute, Chinese Academy of Medical Sciences & Peking Union Medical College, Peking University, Beijing National Laboratory for Molecular Sciences, Novo Nordisk Foundation, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences

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