Fig. 1: Identification of tyrosine phosphorylation as potential regulation of RIPK1 activity.
From: Tyrosine phosphorylation regulates RIPK1 activity to limit cell death and inflammation
HEK293T cells were co-transfected with expression vectors for Flag-RIPK1 and HA-JAK1 (a) or Flag-SRC and HA-RIPK1 (b) for 24 h. Cell lysates were then immunoprecipitated with anti-Flag-Protein A/G agarose and analyzed by immunoblotting with the indicated antibodies. c Primary WT BMDMs were stimulated by Flag-TNF (100 ng/ml) at indicated time points and TNF-RSC was immunoprecipitated by anti-Flag resin for western blotting with indicated antibodies. Primary WT BMDMs were stimulated by TNF (d) or TNF/BV-6 (e) at indicated time points and whole-cell lysates were immunoprecipitated using anti-JAK1 (d) or anti-RIPK1 (e) antibody for immunoblotting with the indicated antibodies. HEK293T cells were co-transfected with expression vectors for Flag-RIPK1 and HA-JAK1 (f) or HA-SRC (g) for 24 h. Cell lysates were immunoprecipitated with anti-Flag-Protein A/G agarose analyzed by immunoblotting with phosphor-tyrosine antibody and other antibodies. h Primary WT BMDMs were stimulated by TNF for 15 min or TNF/BV-6 for 1 h with or without JAK1 and SRC inhibitor treatment. Cell lysates were then harvested and immunoprecipitated using anti-phospho-Tyrosine antibody for immunoblotting with the indicated antibodies. TNF: 10 ng/ml; BV-6: 2.5 uM. Source data are provided with this paper.
