Fig. 5.

PTK6 downregulation activates p38 MAPK. a Lysates from MCF-7L-TamR or T47D-TamR cells expressing control or PTK6 shRNAs (C9 and 49) were probed with the indicated antibodies. b MCF-7L-TamR cells were lysed 48 h after shRNA lentiviral infection. Active, phosphorylated p38 was immunoprecipitated and in vitro kinase assay was performed in the presence of ATP (200 μM) and GST-ATF2 substrate (1 μg). The kinase reaction mixture was probed with antibodies to phospho-ATF2, ATF2, phospho-p38 and P38. c Cells expressing either control or PTK6 shRNAs (49 or C9) were treated with DMSO or SB203580 (5 μM). Cells were lysed at 36 and 56 h (MCF-7L-TamR) or 60 and 72 h (T47D-TamR) after shRNA lentiviral infection. Lysates were probed with the indicated antibodies. All experiments were performed three times. All the western blot images were processed in parallel