CAMK2G is phosphorylated by MPL signal via JAK2. (A) Immunoblotting of phospho-CAMK2G in 32D cells overexpressing wild-type MPL together with CAMK2G when cells were stimulated by TPO in the absence of IL-3. β-Actin was used as a loading control. (B) The result of the immunoblotting is quantified. The ratio of phospho-CAMK2G/CAMK2G is shown. N = 4, independent experiments, unpaired 2-tailed Student t test: *P < .05. (C) Immunoblotting of phospho-CAMK2G in 32D cells overexpressing wild-type MPL or MPL W515L together with CAMK2G when cells were treated with ruxolitinib. β-Actin was used as a loading control. (D) Immunoblotting of phospho-CAMK2G in 32D cells overexpressing MPL W515L and CAMK2G after knockdown of JAK2 with shRNA. β-Actin was used as a loading control. (E) Immunoblotting of phospho-CAMK2G in JAK2 knockdown cells after cells were transduced with empty or wild-type JAK2 vectors. β-Actin was used as a loading control. (F) Immunostaining using anti-CAMK2G antibody was performed in Ba/F3_MPLwt and Ba/F3_MPLmu. Scale bar, 10 μm. (G) Immunoblotting of phospho-JAK2, phosphor-STAT5 in Ba/F3_MPLmu when treated with berbamine. β-Actin was used as a loading control.