Fig. 4.

Coimmunoprecipitation of HA-tagged PLM and NKA. (A) Representative immunoblots of the starting material (pre-IP) and the immunoprecipitate (using an agarose-conjugated HA antibody). Recombinant NKA and PLM were detected using GFP and PLM antibodies, respectively. Comparable amounts of PLM were immunoprecipitated for each lane, but variable amounts of NKA were in co-IP, with or without forskolin preincubation. M, molecular weight ladder. (B) Mean data (NKA/PLM signal normalized to control) from different blots (n = 3) show that FRET-abolishing alanine mutations resulted in a reduction, but not an abolition of associated CFP–NKA-α1. *P < 0.01, statistical significance versus control using one-way ANOVA. ^#P < 0.05, statistical significance between pairs (− vs. + forskolin) using Student’s t test.