Fig. 7: p38MAPK phosphorylates the 9th threonine of the CABIN1 peptide. | Experimental & Molecular Medicine

Fig. 7: p38MAPK phosphorylates the 9th threonine of the CABIN1 peptide.

From: The optimized core peptide derived from CABIN1 efficiently inhibits calcineurin-mediated T-cell activation

Fig. 7

a Jurkat T cells expressing HA-mCherry-tagged CABIN1 peptide were treated with 40 nM PMA in a time-dependent manner. The phosphorylated threonine of the immunoprecipitated peptide with HA antibody was detected with [PXpTP] antibody. b In vitro kinase assays using purified GST-tagged CABIN1 peptide and PKC downstream kinases. c In vitro kinase assays using purified GST-tagged peptides and kinases included in the MAPK family. d Immunoprecipitation assays using Flag-MAPKs and HA-CABIN1 in HEK293T cells. e, f In vitro kinase assays using p38α and substituted CABIN1 peptides. g Jurkat T cells expressing HA-mCherry-tagged CABIN1 peptide were treated with 10 μM p38α inhibitors for 16 h. Immunoprecipitation was performed with HA antibody and detected by western blotting. h Phosphorylated threonine and peptide-bound CNA of g were measured using ImageJ, and the ratio of phosphorylation and calcineurin binding to the CABIN1 peptide was calculated (n = 6). Values shown are the mean ± standard deviation. Statistical differences were determined using one-way ANOVA. *p < 0.05, **p < 0.01, ***p < 0.001, CBB Coomassie Brilliant Blue, NC negative control.

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