Figure S11 from ERBB4-Mediated Signaling Is a Mediator of Resistance to PI3K and BTK Inhibitors in B-cell Lymphoid Neoplasms

Protein levels of ERBB4 was measured by FACS (A) and immunoblotting (B) in JEKO1 cells engineered to increase the expression of ERBB. Drug sensitivity for idelalisib (C) and ibrutinib (D) was evaluated in the different clones of engineered JEKO1. (A) Representative plot of two independent experiments. Boxplot and density plot shows the median MFI values of the two replicates: negative control (black), JEKO1-CNT (control, grey) and JEKO1-ERBB4 (sgRNA-erbb4, red). Results are presented from Welch Two Sample test performed on MFI values of the two experiments. (B) Immunoblotting for ERBB4 in JEKO1-CNT (control, grey) and JEKO1-ERBB4 (sgRNA-erbb4, red). Representative image of two independent experiments. Data on the barplot represents the mean of two experiments, errors bars represent standard deviation of the mean. * for p<0.05 from a t-test. (C-D) Cell viability was evaluated by MTT assay (72h). JEKO1-CNT (control, blue) and JEKO1-ERBB4 (ERBB4-sgRNA3 in red) cells were stimulated with PBS (left panel) or HBEGF (30ng/mL, right panel) for 4 hours and the exposed to increasing concentrations of idelalisib (C) or ibrutinib (D). Error bars correspond to standard deviation of the mean. Data derived from two independent experiments. Statistically significant differences were evaluated by moderated t-test comparing each treatment to control. * for p<0.05.