Expression of WNT11 protein by QCE6 cells and WNT11 stable transfectants.

Each lane contains protein isolated from cultures of WNT11ox/3 (lane 1), QCE6 (lane 2), WNT11αs/6 (lane 3), or WNT11αs/4 cells (lane 4). Confluent cultures of each cell line were treated overnight with 50 μg/mL heparin (which solubilizes WNT protein bound to cell surfaces and matrix). After culture fluids were collected, total cell protein was isolated following cell lysis in hypotonic buffer. The top panel shows cell supernatants blotted for WNT11, as indicated by an individual band of approximately 48 kd (arrow). The greatest amount of WNT11 protein was exhibited by WNT11ox/3 cells (lane 1), with QCE6 cells showing more moderate levels of this protein (lane 2). In comparison, both WNT11αs/6 (lane 3) and WNT11αs/4 cells (lane 4) produced very low amounts of WNT11. The bottom panel shows total cell protein (5 μg/lane) blotted for pyruvate carboxylase to ensure that the volumes of culture supernatants added to the WNT11 gel were representative of similar cell numbers.