QUICK SEARCH:   [advanced] Author: Keyword(s):
Home     Help     Feedback     Subscriptions     Archive     Search     Table of Contents

Help viewing high resolution images Return to article

Fig. 4. Phosphorylation of PTPN13 by PKA in vitro and in vivo. (A) HEK293 cell lysate was subjected to immunoprecipitation using control IgG (lane 1-4) or anti-human PTPN13 antibodies (lane 5-12). Immunoprecipitates were then incubated with or without PKA catalytic subunits for the indicated time followed by 6% SDS-PAGE analysis. Radiolabeled PTPN13 was detected by autoradiography. (B) [³²P]-metabolic-labeled HEK293 cells were stimulated with 100 µM forskolin for the indicated time. Harvested cell lysates were then subjected to immunoprecipitation using an anti-human PTPN13 antibody or control IgG, followed by 6% SDS-PAGE analysis. Radiolabeled and total PTPN13 were detected by autoradiography (upper panel) or western blotting analysis (lower panel). Densitometric analysis was performed on four independent experiments and the average and SEM are reported in the bar graph. (C) Human PTPN13 was immunoprecipitated by anti-PTPN13 antibody or normal rabbit IgG from HEK293 cell lysates. Immunoprecipitated PTPN13 was then subjected to in vitro phosphorylation with or without PKA catalytic subunit (PKA), PKA + PKI, or heat-inactivated PKA (HI-PKA). After washing three times with lysis buffered solution, tyrosine phosphatase activity in the immunoprecipitates was measured.

Return to article