TMP decreases BTK expression and enhances growth inhibitory effect of ibrutinib against WM cells. (A) BCWM1 cells with or without TMP treatment at various concentrations for 24 hours were subjected to WB analysis using anti-BTK and GAPDH. (B) BCWM1 (left) and MWCL1 cells (right) were cultured in the presence of different doses of ibrutinib and TMP alone, or in combination for 24 hours. Cell growth was assessed by [³H]thymidine uptake and presented as a percentage of control cells (untreated cells). Data are means ± SD (n = 3). (C) CIs and fractions affected are shown in the graphs (upper panel) and in the tables (lower panel) for BCWM1 (left) and MWCL1 (right) cell lines. (D) Caspase 3/7 activity was evaluated in BCWM1 and MWCL1 after single and combination drug treatment using Caspase-Glo 3/7 assay. (E) Total and phosphorylated STAT3 levels in cell lysates from cells treated with or without TMP and ibrutinib alone or in combination were evaluated with STAT3 (Total/Phospho) InstantOne ELISA. (F) BCWM1 cells were treated with TMP (10 µM), 407601 peptide (10 µM), ibrutinib (1 µM), or combined therapy for 48 hours, followed by Annexin V/propidium iodide staining and flow cytometry analysis.