Phosphorylation of JAK2 and Lyn.

CD34⁺ cells stimulated with EPO for 4 days were again incubated with or without the addition of NU-L_PAN for 18 hours. Incubated cells were restimulated with 5 U/mL EPO for 0, 5, and 15 minutes, and then cell lysates were prepared. JAK2 (A), phosphorylated JAK2 (B), Lyn (C), and phosphorylated Lyn (D) were detected by Western blotting using anti-Jak2, antiphosphorylated JAK2, anti-Lyn and antiphosphorylated tyrosine antibodies, respectively. For the detection of phosphorylated Lyn bands by antiphosphorylated tyrosine antibody, the bands corresponding to the molecular weight of Lyn were inferred to as phosphorylated Lyn. A result representative of 3 separate experiments is presented.