Extended Data Fig. 3: Validation of 2dV-Camui conformational CaMKII FRET sensor with wildtype, T286A or T305D/T306D CaMKII mutants.
From: Dendritic, delayed, stochastic CaMKII activation in behavioural time scale plasticity
a, Grayscale and fluorescence lifetime images of HeLa cells expressing 2dV-Camuiα sensors, which have wildtype CaMKII (WT), CaMKII mutant deficient autonomous activation (T286A), or CaMKII mutant deficient in Ca2 + /calmodulin binding (T305/T306D). Each image is before (pre), 1 min after ionomycin (3 µM) and 2.5 min after EGTA (8 mM) application. b, c, Averaged time course (b) and summary of the peak (1 min post-ionomycin application) (c) of fluorescent lifetime changes of 2dV-Camui in WT and mutant CaMKII expressing HeLa cells in response to bath application of ionomycin. The data are presented as mean ± s.e.m. (n = # of cells/# of independent experiments: WT, n = 91/8; T286, n = 63/6; T305D/T306D, n = 74/6). c, One-way ANOVA (F(2, 225) = 265.5) followed by Dunnett’s post hoc test (WT vs. T286A, p = 4.0e-13; WT vs. T305D/T306D, p = 4.0e-13). ****p < 0.0001.
