Fig. 1
a Fragments Per Kilobase Million (FPKM) of TET1, TET2, TET3 in a cohort with 124T-ALL patients and 12 normal T cell samples (CNCB, HRA000122). Data are mean ± SD (Two-tailed unpaired Student’s t test, ** p < 0.01, *** p < 0.001). b Transcripts per million (TPM) of TET1 in glucocorticoid-resistant and glucocorticoid-sensitive T-ALL samples (GSE5820). c Protein expression levels of TET1, TET2, TET3 in 8 T-ALL cell lines and normal T cells. d Effect of TET1 knock down on cell growth of Jurkat cell line. e Schematic of structure guided virtual screening and cell line-based drug screening with 2059 FDA or EMA approved drugs. Jurkat and CCRF-CEM cell lines were treated with each drug (5 μM) for 48 h, and cell viability was detected using CCK-8 assay. f Joint analysis of virtual screening and cell line-based screening identified auranofin as a potential hit drug. g Affinity between TET1 protein and auranofin assayed by surface plasmon resonance (SPR). Equilibrium binding analysis indicates a KD of 1.804 μM. h Dot blot analysis of global 5hmC in Jurkat cell showing that auranofin dose-dependently decreased cellular 5hmC level. Left: dot blot image; right: quantification of dot blot results. i Quantification of genome-wide 5hmC distribution in T-ALL Jurkat cells treated with or without 0.1 μM auranofin for 24 h. Auranofin treatment decreased global 5hmC levels. n = 5 replicates. j Dose-dependent inhibition of TET1 catalytic activity by auranofin with an in vitro fluorometric quantification assay. IC50 of 0.076 μM was determined, indicating potent inhibition of TET1 by auranofin. k Molecular docking showing that auranofin are in close conformation with 2-OG and Fe (II) in the binding pocket of TET1-CD. l SPR-based assay showing that the binding of auranofin to TET1 was competed by increasing concentration of TET1 substrate analogue NOG. m Dot blot analysis of 5hmC revealed that auranofin induced TET1 catalytic activity inhibition was attenuated by increasing concentration of 2-OG. Left: dot blot image; right: quantification of dot blot results