Figure 1.

The P1aABD and CaM Interact In Vivo

(A) 804G cells ectopically expressing combinations of P1aABD-CFP and P1aABD_mut-CFP with CaM-YFP or YFP alone. FRET was measured by acceptor photobleaching. Pseudocolor pre- and postbleach images of acceptor (CaM-YFP or YFP alone) and FRET images (FRET_eff) are shown. FRET efficiency was determined as a relative increase of donor fluorescence in the cytoplasmic region of interest shown in boxed area, and enlarged in insets (bottom right corners). Bar, 10 μm.

(B) Box and whisker plots indicate the median FRET (middle line in the box), 25th percentile (bottom line of the box), 75th percentile (top line of the box), and minimum and maximum values (whiskers). A CFP-YFP fusion was used as a positive control for FRET (first column); cotransfection of P1aABD-CFP and empty YFP vector was used as a negative control (last column). The number of assessed cells/individual bleach events (n) obtained from three or more independent experiments is indicated.