In vitro effect of combined therapy with dasatinib and BTKi on primary ALL samples. (A) Flow cytometric analysis of primary samples aiming to detect the phosphorylation status of PLCG2 and BTK after 30 minutes of treatment with vehicle (DMSO), dasatinib, ibrutinib, and dasatinib + ibrutinib. Gating strategy on lymphocytes, single cells, 7AAD, CD19⁺, and CD117⁺. One representative histogram per antibody is showed. (B-C) Histogram represents the phosphorylation of (B) PLCG2 and (C) BTK in primary ALL samples with different karyotypes, after treatment with vehicle, ibrutinib, dasatinib, and dasatinib + ibrutinib treatment.