Fig. 8.

Effects of reduced CaM binding at individual SMDs on AngII-induced Ca²⁺ signals. HEK293 cells expressing plasmids encoding mKate2 alone or in fusion with the N terminus of full-length AT₁R with wild-type or mutant sequence at SMD2, or SMD4_JM were loaded with fura-2/AM as described under Materials and Methods. (A) Epifluorescence of mkate2 moiety. (B) Corresponding epifluorescence of fura-2 in the same microscopic field. (C) Merged mKate2 and fura-2 fluorescence images. (D) Average mKate2 fluorescence intensities of all individual cells selected for Ca²⁺ measurements. (E) Average time courses of Ca²⁺ signals stimulated by AngII as indicated. (F) Corresponding average total integrated areas under curve (AUC). N = 60 cells from 6 independent experiments for each paradigm; * and ^ϕ, p < 0.05 vs AUC values of cells expressing mKate2 only and mKate2-wildtype AT₁R fusion, respectively.