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. 2020 Oct 23;35(6):1621–1630. doi: 10.1038/s41375-020-01055-7

Fig. 4. Wnt5a induces ROR1-dependent phosphorylation of DOCK2 in CLL.

Fig. 4

a Tyrosine phosphorylation of DOCK2 was confirmed by immunoblot analysis of anti-DOCK2 immune precipitates (ip), using lysates of MEC1-Ctrl, MEC1-ΔPRD, MEC1-ROR1 (W/T) or MEC1 cells transfected with each of the various mutated forms of ROR1, as indicated on the top; the membranes were probed with anti-DOCK2 or anti-phospho tyrosine antibody (pDOCK2), as indicated on the left. b Immunoblot analysis of anti-DOCK2 ip, using lysates prepared from MEC1-ROR1 cells that had been treated with Wnt5a neutralizing antibody (2 µg/ml, R & D) for the times indicated at the top (in hours); membranes were probed with anti-phospho tyrosine (pDOCK2) or anti-DOCK2 antibody, as indicated on the left. c Immunoblot analysis of anti-DOCK2 ip, using lysates prepared from MEC1-ROR1 cells that had been treated with cirmtuzumab (20 µg/ml) for the times indicated at the top (in hours); membranes were probed with anti-phospho tyrosine (pDOCK2), anti-DOCK2 antibody, as indicated on the left. d Immunoblot analysis of anti-DOCK2 ip, using lysates prepared from overnight, serum-starved primary CLL cells (representative of 3 patients) that subsequently were treated for 5 min without (-) or with (+) Wnt5a (100 ng/ml), as indicated on the top; the membranes were probed with anti-DOCK2 or anti-phospho tyrosine antibody (pDOCK2), as indicated on the left. e Immunoblot analysis of anti-DOCK2 ip, using lysates prepared from overnight, serum-starved primary CLL cells (representative of 3 patients) that subsequently were treated with Ctrl-IgG or cirmtuzumab (20 μg/ml), without (-) or with (+) Wnt5a (100 ng/ml), as indicated on the top; the membranes were probed with anti-DOCK2 or anti-phospho tyrosine antibody (pDOCK2), as indicated on the left.