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. Author manuscript; available in PMC: 2013 Aug 15.
Published in final edited form as: Dev Biol. 2012 Jun 7;368(2):335–344. doi: 10.1016/j.ydbio.2012.05.036

Figure 3. ADAM13 function is dispensable for CNC migration in vitro.

Figure 3

(A) ADAM13 and 19 knock down has no effect on CNC migration in vitro. Embryos were injected in one cell at the two-cell stage with a mixture of GFP mRNA and either control morpholino (Ctl), ADAM13 morpholino (MO13) or ADAM13 and 19 morpholino (2MO). At stage 15, embryos were sorted and CNC explants placed on fibronectin-coated 96-well plates. For each case, pictures of typical explants are shown before migration (t=0), at the end of the sheet migration phase (t=6h) and at the end of the single cell migration phase (t=16h). Morphant CNC cells were capable of migrating in vitro in a pattern indistinguishable form the control CNC. (B) ADAM13 knock down in CNC is efficient and prevents Cadherin-11 cleavage. Twenty CNC were dissected at stage 15 and placed on FN substrate for 2 hours before cell surface proteins were biotinylated. Proteins were extracted and immunoprecipitated sequentially using a goat anti-ADAM13 antibody (gA13), the mouse mAb to Cadherin-11 (1B4) and the mouse mAb to integrin 1 (8C8). Both gA13 and 1B4 recognize the cytodomains of ADAM13 and Cadherin-11, respectively. ADAM13 knock down completely abolishes the expression mature ADAM13 at the cell surface (ADAM13, arrowhead). The full length Cadherin-11 protein is still strongly expressed (Cad-11, arrowhead a) but is no longer cleaved (Cad-11, arrowhead b). Integrin α5β1 cell-surface expression is unaffected by ADAM13 knockdown.