<p><i>A</i>, T305 phosphorylation was detected by Western analysis after reactions corresponding to the kinase activity assays shown in Fig. 1 (1 min at 30°C, but without substrate peptide) or after extended reaction times (10 min). <i>B</i>, Quantification of the relative T305 phosphorylation by the ratio of the phospho-T305 and total CaMKII immuno-detection values (IDV) shows significantly lower phosphorylation of the T286A mutant in our 1 min kinase assay conditions (N = 3 different extracts for each kinase form; *: p<0.05 in two-tailed t-test). The IDV ratio for T286A was ∼28% (±7.6%) of wild type after the 1 min reactions; in the 10 min reaction experiment shown in panel A, this ratio was ∼83%, consistent with the faster wild type reaction reaching saturation faster.</p
