Figure 1.
14-3-3 isoforms interact with TET2. A, peptide counts from LC-MS analysis of immunoprecipitates from WT and S99A TET2 (N-terminal 400 amino acids). Analysis of control IgG control is also included. B and C, Myc-TET2 WT and Myc-TET2 S99A were immunoprecipitated from HEK293T cells transiently transfected with the respective plasmids (8 μg of WT TET2, 10 μg of S99A TET2) and analyzed by IB for 14-3-3-θ (B) and 14-3-3-ϵ isoforms (C). Whole-cell extracts were used as inputs. Control IgG and Myc immunoprecipitations were performed from Myc-TET2 and CV-transfected cells, respectively, as negative controls. D, HEK293T cell lysate was immunoprecipitated with anti-TET2 and control IgG antibodies and analyzed by immunoblotting for endogenous 14-3-3 interaction. Data are representative of two independent experiments. E, sequence homology of TET2 around Ser-99 among different species demonstrating a conserved phosphoserine 14-3-3 binding motif ((R/K)XXpSXP). F, WT and S99A TET2 were immunoprecipitated from HEK293T and analyzed by IB with phosphoserine 14-3-3–binding motif antibody.