Figure 5

2D Western Analysis of AKAP7δ and AKAP7γ. Recombinant mouse AKAP7δ, AKAP7γ, and rat AKAP7δ were analyzed by two-dimensional western blotting. V5-tagged Mouse AKAP7δ (A), AKAP7γ (B) or rat AKAP7δ (C) were detected from transfected HEK293 cell lysates using a V5-HRP antibody. (D) Mock-transfected HEK293 cell lysates were probed with V5-HRP as a negative control. (E) Rat heart lysate used in AKAP7 immunoprecipitations (using AKAP7 antibody) was probed with protein A-HRP. Two products are identified, rat AKAP7δ, and a variant of rat AKAP7δ identified in this study (rat AKAP7δ variant 1, see Supplemental Information Figure 3). (F) Mouse heart lysate subjected to AKAP7 immunoprecipitation followed by detection with protein A-HRP. Mouse AKAP7δ and AKAP7γ are identified in addition to two unknown splice variants (indicated by red arrows). For negative control immunoprecipitations rat heart lysate (G) or mouse heart lysate (H) was immunoprecipitated with rabbit IgG (pre-immune serum) followed by detection by protein A-HRP. The pH range and molecular weight are indicated at the top and left (respectively) of each blot.