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. 2017 Jun 16;31(12):2615–2622. doi: 10.1038/leu.2017.133

Figure 4.

Figure 4

HS1 associates with ARHGEF1, which undergoes HS1-dependent activation to enhance activation of RhoA. (a) Immunoblot analysis of anti-HS1 ip or Ctrl-IgG ip, as indicated at the top, using lysates prepared from freshly isolated primary CLL cells; the membranes were probed with anti-HS1 or anti-ARHGEF1 antibody, as indicated on the left. (b) Immunoblot analysis of anti-ARHGEF1 ip or Ctrl-IgG ip, as indicated at the top, using lysates prepared from freshly isolated primary CLL cells; membranes were probed with anti-HS1 or anti-ARHGEF1 antibody, as indicated on the left. (c) In vitro exchange assay on RhoA of anti-ARHGEF1 ip from lysates of CLL cells transfected with Ctrl-siRNA (green line) or siRNA specific for HS1 (red line) in the presence of Wnt5a. The green line depicts GTPase-activation using buffer alone. (d) Immunoblot analysis of lysates prepared from MEC1-ROR1 cells transfected 72 h previously with control siRNA or siRNA targeting HS1; expression of HS1, total RhoA, and activated RhoA was measured, as indicated on the left.