Gα12QL activates JNK in breast cancer cells.

<p>MDA-MB-231 cells were infected with GFP (control) or Gα12QL adenovirus for 6–7 h. (A) Cells were starved for 2 h and then treated for the indicated time with JNK inhibitor SP600125 (20 µM) prior to lysis in JNK activation buffer and processing for analysis (see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0026085#s2" target="_blank"><i>Materials and Methods</i></a>). (B) Cells were transfected with JNK siRNA or control (luc) siRNA for ∼72 h prior to infection with adenovirus as in (A). Cells were starved for 18 h before being lysed. For both panels, equal amounts of total protein in lysates were subjected to SDS-PAGE and immunoblot analysis to assess levels of phospho-JNK (P-JNK, p54 and p46) and phospho-c-Jun (P-c-Jun). Total levels of JNK and Gα12 expression, along with α-tubulin as a loading control, are also shown. Each panel shows the results of a single experiment that is representative of three or more independent experiments.