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. 2018 Dec 3;8:560. doi: 10.3389/fonc.2018.00560

Figure 1.

Figure 1

Both JH1 and JH2 in JAK1 and JAK3 are required for functional IL-2 signaling. (A) Schematic presentation of JAK1 and JAK3 wild-type (WT) and domain deletion constructs. (B) Both JH1 and JH2 of JAK1 and JAK3 are required for IL-2 dependent STAT5 activation. U4Cγβ cells were transfected with JAK1 WT, JAK3 WT and domain deletion constructs together with STAT5 responsive reporter SPI-luc2 (28), either with Renilla or pRL-TK control vector (see Materials and Methods) and starved/stimulated. Similar results were obtained with both control vectors. Data shows averages and SD from normalized values combined from three independent experiments, each having triplicate samples. The expression of JAK1/JAK3 constructs was analyzed by anti-HA Western blotting. (C) IL-2-induced pSTAT5 requires both JH1 and JH2 in JAK1 and JAK3. STAT5 phosphorylation and expression of JAK1, JAK3 and STAT5 were detected from cell lysates from transfected U4Cγβ cells by immunoblotting with HA- and pTyr694 STAT5 antibodies. Similar results were obtained in three independent experiments.