Inhibition of miR-17∼92 blunts the BCR response in DLBCL cell lines. (A-C) The DLBCL cell lines HBL-1, TMD8, U2932, OCI-LY10, OCI-LY1, SUDHL4, Pfeiffer, and Karpas 422 were treated with either control short-hairpin inhibitor or miR-17∼92 short-hairpin inhibitor mix for 48 hours. (A) The DLBCL cell lines were harvested and western blotting was performed for CD22 (two exposures shown), FCGR2B, FCRL4, and actin. (B) The DLBCL cell lines were treated with treated with 5 μg/mL soluble anti-IgM and anti-IgG or 10 μg/mL soluble isotype control for 15 minutes. Western blotting was performed for P-SYK, total-SYK, P-BLNK, total-BLNK, and actin. (C-D) The average (mean) normalized Myc (C) and mir17hg (D) expression in the various DLBCL subtypes classified by the consensus signature. Error bars represent the SEM. One-way analysis of variance P values are denoted. (E) GSA demonstrating a positive correlation between Shipp BCR signature genes and averaged mature miR-17∼92 expression. The heatmap shows the expression of the 41 genes in columns with each tumor sample in rows. Orange indicates high expression; blue indicates low expression. For each gene, the gene score from the GSA is indicated in the plot above the heatmap, with positive scores indicating positive association with miR-17∼92 expression. The averaged mature miR-17∼92 expression for each sample is shown in the graph to the right of the heatmap.