Extended Data Fig. 2: Phosphorylation regulation of THEMIS at Tyr34 site.
From: THEMIS is a substrate and allosteric activator of SHP1, playing dual roles during T cell development
a, Sequence homology comparison of THEMIS Tyr34 site in 9 species. b, THEMIS amino acid sequence showed a 31aa-long peptide after trypsin digestion in the red box. c, As indicated, HEK293T cells were transiently transfected with THEMIS-Myc and GRB2-FLAG or LCK-FLAG. Co-immunoprecipitation assay demonstrated that the interaction between LCK and THEMIS was transient and weak compared to GRB2. d-e, Immunoblotting of truncation variant experiments in the HEK293T cell line exploring the three domains of THEMIS with five different truncation mutants (d), Western blotting results showing that LCK can only phosphorylate the THEMIS Tyr34 site, positioned in the CABIT1 ___domain (e). f, Sequence alignment between hGARE1, mGARE1, hGARE2, mGARE2, mTHEMIS3, hTHEMIS1, mTHEMIS1, hTHEMIS2, and mTHEMIS2, showing one conserved Tyr residue (Y34 in THEMIS and Y35 in THEMIS2, respectively).
