Tyrosine phosphorylation of gp130, Jak 2, and STAT3 in murine calvaria cells treated with IL-11 and/or heparin.

Confluent calvaria cells were stimulated for 10 minutes with either heparin alone (25 μg/mL), IL-11 alone (20 ng/mL), or heparin plus IL-11 (25 μg/mL and 20 ng/mL, respectively). (A) The cells were lysed and the lysate was immunoprecipitated with a polyclonal rabbit antimouse gp130 antibody. Precipitates were immunoblotted with an anti-gp130 antibody (upper panel) or run on a second gel before immunoblotting with a monoclonal anti-pTyr antibody (lower panel). (B) Cell lysates were immunoprecipitated with a polyclonal rabbit antimouse Jak2 antibody prior to immunoblotting the precipitates with an anti-Jak2 antibody (upper panel) or loaded on a second gel and immunoblotted with a monoclonal anti-pTyr antibody (lower panel). (C) Cell lysates were immunoprecipitated with a polyclonal rabbit antimouse STAT3 antibody prior to immunoblotting the precipitates with an polyclonal anti-STAT3 antibody (upper panel) or running them on a second gel and immunoblotting with a polyclonal pTyr-STAT3 antibody (lower panel).