Wnt5a induces ROR1-dependent production of proinflammatory factors through activation of NF-κB in CLL cells. (A) Immunoblot analysis of lysates from CLL cells treated by Wnt5a (200 ng/mL) for 30 minutes, with or without cirmtuzumab, and probed using anti-phospho-p65, anti-p65, anti-phospho-IκBα, anti-IκBα or anti-β actin. (B) Immunoblot analysis of lysates from CLL cells cocultured with NLC overnight, with or without cirmtuzumab or anti-Wnt5a, and probed with anti-phospho-p65 or anti-p65. (C) ELISA of IL-6 in medium of CLL cells cultured with Wnt5a and BAY 11-7082 (5-20 µM) for 8 hours, as indicated in the text. (D) Immunoblot analysis of lysates from CLL cells treated by Wnt5a and BAY 11-7082 (5-20 µM) for 8 hours that was probed with anti-pSTAT3 (Y705) or anti-STAT3, as indicated in the text. (E) Volcano plot of transcripts of NF-κB target genes expressed by CLL cells stimulated with Wnt5a in the presence of control IgG or cirmtuzumab (P < .05 was considered statistically significant). (F) Heat map of transcripts of NF-κB target genes expressed by CLL cells stimulated with Wnt5a in the presence of control IgG or cirmtuzumab.