Extended Data Fig. 2: The enzymatic activities of LCKWT and LCKCA in cells are comparable.
From: Unique roles of co-receptor-bound LCK in helper and cytotoxic T cells
(a) HEK293 cells were co-transfected with LCK-FLAG variants and CD25-TCRζ fusion protein or empty vector (e.v.) and harvested after 24 h. The phosphorylation of CD25-TCRζ was by detected immunoblotting. A representative experiment out of 2 in total. (b) HEK293 cells were co-transfected with LCK-FLAG variants and ZAP70 or empty vector (e.v.) and harvested after 24 h. The phosphorylation of ZAP70 was by detected immunoblotting. A representative experiment out of 2 in total. (c) LCKKO Jurkat cells reconstituted with indicated LCK variants were activated by anti-TCR antibody for 90 s. The phosphorylation and expression of indicated markers was detected by immunoblotting. A representative experiment out of 3 independent experiments. (d) LCKKO Jurkat cells expressing OT-I TCR were reconstituted with various LCK variants and activated with OVA-loaded T2-Kb cells overnight and analyzed for CD69 expression by flow cytometry. Mean + /- s.e.m. is shown. n = 3 independent experiments.
