Journal: The Journal of Biological Chemistry

Article Title: The stress-activated kinase p38 mediates non-canonical activation of Src and tyrosine phosphorylation of the adapter protein TAB1

doi: 10.1016/j.jbc.2026.111200

Figure Lengend Snippet: Src directly phosphorylates TAB1-Y481. A , the domain structure of TAB1 and the amino acid sequence around Y481. Conserved phosphorylation serine/threonine and tyrosine sites in the C-terminal regions are highlighted in green / blue and red . B , COS-7 cells were transfected with EGFP-tagged full-length (FL), N-terminal (N), or C-terminal (C) TAB1 and Src. C , COS-7 cells were transfected with EGFP-TAB1 and Src. Dasatinib (0.1 μM) was added 1 h before cells were harvested. D , COS-7 cells were transfected with TAB1-C or its Y481F mutant (YF) and Src. Cell lysates and immunoprecipitates with an anti-GFP antibody were immunoblotted with anti-phosphotyrosine and anti-GFP antibodies. The expression of active Src was detected by an anti-phospho-Src family kinase (Y419) antibody ( B – D ). E , an in vitro kinase assay was performed using recombinant GST-TAB1-C and active GST-Src proteins. F , COS-7 cells were transfected with EGFP-TAB1 with SFKs, including Src, Fyn, Lyn, and Lck. G , HCT-116-Src cells were treated with 10 ng/ml doxycycline (Dox) for 24 h. TAB1-Y481 phosphorylation and other proteins were detected by an anti-phospho-Y481-TAB1 antibody and the antibodies described above, respectively ( E – G ). H , the relative quantification of pY481-TAB1, normalized to total TAB1, is presented as the mean ± SD from three independent experiments. p values were calculated using Welch’s two-tailed t test. ∗ p < 0.05.

Article Snippet: The recombinant human GST-TAB1-C protein (WT and Y481F) derived from Escherichia coli ( ) was reacted with the recombinant human active GST-Src kinase derived from insect cells (Carna Bioscience) at 30 °C for 30 min in 30 μl of reaction buffer containing 20 mM HEPES (pH 7.6), 20 mM MgCl 2 , 0.2 mM ATP, 2 mM DTT, 20 mM β-glycerophosphate, and 0.1 mM sodium orthovanadate.

Techniques: Sequencing, Phospho-proteomics, Transfection, Mutagenesis, Expressing, In Vitro, Kinase Assay, Recombinant, Quantitative Proteomics, Two Tailed Test

Journal: bioRxiv

Article Title: Slap restricts oncogenic Src-family kinase signaling to maintain colonic epithelial homeostasis

doi: 10.64898/2026.01.05.697659

Figure Lengend Snippet: (A) Slap-dependent colonic organoid development. Representative images of colon-derived organoids from Slap f/f and Slap f/f Villin-Cre ERT2 mice cultured in Matrigel for 2 days. Quantification of organoid number and area is shown as mean ± SEM from 100-200 organoids per mouse (n = 8 mice per group). ****p<0.0001 Mann–Whitney test. (B) Intestinal Slap deletion increases SFK activity and global protein tyrosine phosphorylation in colonic crypts. Representative immunoblots (right) and quantification (left) of phospho-SFK (pSRC) and total phospho-tyrosine levels in lysates from isolated colonic crypts of the indicated genotypes. Data are presented as mean ± SEM from n = 4-5 mice per group. *p<0.05; **p<0.001 t test. (C) Slap-dependent colonic organoid expansion requires SFK activity. Representative images and quantification of colon-derived organoids from Slap f/f and Slap f/f Villin-Cre ERT2 mice cultured for 2 days in the presence or absence of the SRC-family kinase inhibitor eCF506 (100 nM). Data are shown as mean ± SEM from 100-200 organoids per mouse, n = 4 mice per group. ***p<0.001, ****p<0.0001 Mann–Whitney test.

Article Snippet: After Matrigel polymerization, 500 μL of M2 medium (M1 media supplemented with EGF (50 ng/mL, Bio-techne), Noggin (100 ng/mL, Stem cell technologies), R-spondin1 (500 ng/mL, Stem cell technologies), Y27 (10 μM, Sigma-Aldrich), CHIR-99021 (3 μM, Tebu-Bio) and WNT3a (50 n/ mL, Thermo Fisher Scientific) was added to each well containing or not the SRC inhibitor eCF506 (100 nM, Medchem express) and in other cases the EphB inhibitors EPHB2i (ALW-II-49-7, 200 nM) and the pan-EPH inhibitor pan-EPHi (ALW-II-41-27, 100 nM, Medchem express).

Techniques: Derivative Assay, Cell Culture, MANN-WHITNEY, Activity Assay, Phospho-proteomics, Western Blot, Isolation