Fig. 5: Single-molecule TIRFM.
From: Hub stability in the calcium calmodulin-dependent protein kinase II
Duplicate experiments (>20 records/experiment), with typically 30–50 immobilized V-CaMKII spots per record, were conducted for each condition (>1000 spots). Spot intensities were averaged over 20 s (100 frames). a The αF394A and βF458A bimodal intensity distributions are relative to the βmonomer and βholoenzyme distributions. The higher mode intensity was double that of the lower mode. The latter superimposed with the mode of the βmonomer distribution (IUNI) The βF458A population fraction with intensity values > 4*IUNI was <10%. The F-test statistic (Fobs/Fpred = 1.22/1.84 < 1) indicates that the variances of the αF394A and βF458A distributions do not differ (probability p > 0.05). b Time-dependent room temperature disassembly in βT287.306-307A holoenzyme populations revealed by comparison of measurements on 30 spots from different slides observed 15 (409 steps) and 45 (384 steps) minutes after dilution from the stock on ice. The F-test statistic (Fobs/Fpred = 0.97/0.39 > 1) indicates there is a significant difference between the two populations (p < 0.05).
