Figure 3. Map7/7D1 bind to Dvl, a key mediator of Wnt5a signaling.

1. Lysates from HeLa cells transfected with control vector or pcDNA3.1‐hMAP7‐V5His₆ were immunoprecipitated with an anti‐V5 antibody, and the immunoprecipitates were probed with anti‐Dvl2 and anti‐V5 antibodies.
2. Lysates from HeLa cells were subjected to immunoprecipitation with control IgG or an anti‐Dvl2 antibody and analyzed by immunoblotting with an anti‐Map7/7D1 or anti‐Dvl2 antibody.
3. Lysates from HeLa cells co‐expressing various deletion derivatives of hMap7‐V5His₆ with EGFP‐mDvl2 were immunoprecipitated with an anti‐GFP antibody, and the immunoprecipitates were probed with anti‐V5 and anti‐GFP antibodies.
4. Lysates from HeLa cells co‐expressing various derivatives of EGFP‐mDvl2 with hMap7‐V5His₆ were immunoprecipitated with an anti‐V5 antibody, and the immunoprecipitates were probed with anti‐GFP and anti‐V5 antibodies.
5. MBP‐hMap7^1‐265ΔCC1 (30 pmol) conjugated to amylose resin was incubated with purified GST, GST‐hDvl1^DIX, or GST‐hDvl1^DEP (30 pmol of each), and the bound proteins were analyzed by immunoblotting with anti‐GST and anti‐MBP antibodies. The positions of GST, GST‐hDvl1^DIX, and GST‐hDvl1^DEP were revealed by loading 1.5 pmol (5%) of each purified protein.
6. Depletion efficiency of siWNT5A or siDVLs. Lysates derived from the indicated cells were probed with anti‐Dvl2 and anti‐Wnt5a antibodies. The blot was reprobed for γ‐tubulin as a loading control (left). Effects of siWNT5A or siDVLs on the protein levels of Map7 and Map7D1 were also analyzed (right). The blot was reprobed for γ‐tubulin as a loading control.
7. Graph shows the relative mRNA levels of MAP7 and MAP7D1 in the indicated cells 72 h after siRNA transfection. Expression levels of MAP7 and MAP7D1 transcripts were quantified by normalization to the GAPDH expression. Data are from three independent experiments and represent average ± SD.

Source data are available online for this figure.