Fig. 2.
CaL association with α5- or β1-integrin depends on adhesion of human embryonic kidney cells (HEK293-T) to fibronectin (FN). A: representative blot showing coimmunoprecipitation (co-IP) protocol on protein lysed from control (untransfected) or neuronal WT-CaL-expressing HEK293-T cells plated onto poly-l-lysine-treated dishes. No α1C-CaL appears to be pulled down by IP with anti-β1-integrin antibody (Ab) in either control (untransfected) or WT CaL-expressing cells (lanes 2 and 7, respectively). Expression of α1C-CaL was confirmed in cells expressing WT-CaL by immunoblotting (IB) with anti-α1C-CaL Ab, showing a specific band at molecular mass 190 to 240 kDa (lane 4). B: representative blot showing pull-down of α1C by anti-α5 or anti-β1-integrin Ab (lanes 4 and 3, respectively) only from cells expressing WT-CaL, not control (untransfected) cells (lanes 7 and 8, respectively), when plated onto FN-coated plates before IP and IB. All blots were repeated at least three times. Prot-G, protein-G.