Fig. 1. Expression of Wnt-5a protein in various cell populations. (A) Wnt-5a-HA fusion protein was detected in the lysates of Wnt-5a-HA-overexpressing cells, using an anti-HA antibody (lane 1), but was not found in the lysates of Wnt-5a antisense (lane 2) or control (lane 3) cells. (B) The content of Wnt-5a protein (Wnt-5a-HA) in Wnt-5a-overexpressing cells detected by an anti-Wnt-5a antibody, which recognises both endogenous and overexpressed Wnt-5a. (C) Examples of Wnt-5a repression in antisense transfectants: lane 1 shows the absence of Wnt-5a protein in a lysate of NIH 3T3 cells (negative control); lane 2, endogenous expression of Wnt-5a protein in a lysate of control cells; lanes 3–7, repression of endogenous Wnt-5a protein in five antisense transfectants (detected with the anti-Wnt-5a antibody), as compared to endogenous levels in control lane 2. (D) Expression of Wnt-5a-HA in lysates of the MCF-7 cells transfected with Wnt-5a (lane 1) and control MCF-7 cells (lane 2). All membranes were subsequently stripped and reprobed with an anti-actin antibody to confirm protein integrity and equal loading.