Analysis of BCR signaling in primary DLBCL patient samples. (A) Classification of 7 DLBCL primary samples based on their expression profile. The heat map shows the expression (z-score) of 8 GCB and 11 ABC genes in the indicated samples. The computed RNA-seq score distinguishes between GCB, ABC DLBCL, or DH. (B,D,F) Representative flow cytometry analysis to determine the phosphorylation of BTK (B, patient 69487), CD19 (D, patient 20954), and GSK3β (F, patient 20954) in purified lymphoma cells stimulated with anti-BCR (orange) or treated with ibrutinib (blue). Unstimulated cells were used as control (gray). (C,E,G) Quantification of the median fluorescence values (MFIs) of pBTK (Tyr223) (C), pCD19 (Tyr 531) (E), and pGSK3β (Ser9) (G) in the indicated patients normalized to the stained, unstimulated control. Data are represented as bar plots corresponding to the mean ± standard deviation (SD) of 3 replicates. Significant changes between stimulated cells (orange bars) and stimulated cells treated with ibrutinib (blue bars) are labeled with an asterisk (uncorrected P ≤ .05). Unlabeled bars indicate not statistically significant changes.