Identification of the critical subtype of CAMK2 in MF. (A) Gene expressions of CAMKs including CAMK1A, 1B, 1G, 1D, 2A, 2B, 2G, 2D, 3, and 4 were assessed by qPCR in BaF/3 and 32D cells overexpressing wild-type MPL and MPL W515L. (B) Immunoblot analysis of JAK, STAT5, CAMK2G, phospho-JAK, phospho-STAT5, and phosphor-CAMK2G. β-Actin was used as a loading control. (C) qPCR data of CAMK2G messenger RNA (mRNA) in BaF/3 cells overexpressing wild-type MPL and MPL W515L transduced with retroviruses carrying vectors with shRNAs or empty vector. Results are means ± SD. N = 3, independent experiments. (D) Proliferation curves of BaF/3 cells overexpressing wild-type MPL and MPL W515L transduced with retroviruses carrying vectors with shRNAs or empty vector. Results are means ± SD. N = 3, independent experiments. (E) Scheme of replating assay using Camk2g wt/− mouse BM. (F) The result of the colony-forming cell capacity of c-kit⁺ cells of wild and Camk2g knockout mice overexpressing MPL W515L. Results are means ± SD. N = 4, independent experiments. (G) Immunoblot analysis of pCAMK2G after treatment with berbamine (CAMK2G inhibitor) in BaF/3_MPLmu. β-Actin was used as a loading control. (H) Scheme of colony-forming cell assay with berbamine treatment. (I) Colony-forming cell assay was performed with BM cells overexpressing wild-type MPL and MPL W515L in the presence of DMSO or berbamine. Results are means ± SD. N = 3, independent experiments. ANOVA test: *P < .05.