Fig. 2: PI(4,5)P2-induced stabilization of PTEN membrane binding.
a–c Representative TIRFM images of membranes composed of 10 mol% (a, c) or 1 mol% (b) PI(4,5)P2 in the absence (a) or presence (b, c) of PTEN-Halo-TMR. Scale bars, 10 μm. d Probability density distribution of the displacement of single molecules in a unit time interval (Δt = 16.5 ms) on membrane containing 1 mol% (magenta), 5 mol% (orange), or 10 mol% (blue) PI(4,5)P2. e PI(4,5)P2 density dependency of the average diffusion coefficient (rectangle) and the diffusion coefficient for state 1 (circle) and state 2 (triangle). Welch’s t test; comparisons were made with “1 mol% PI(4,5)P2” column for the same state. Black, blue, and red indicate D1, D2, and average D, respectively. f Estimation of the number of mobility states using the data obtained from each video. Light, middle, and dark colors indicate the ratio of the videos in which the AIC values show AIC1 − AIC2 < 0 (1 state model), 0 < AIC1 − AIC2 < 4 (2 state model) and 4 < AIC1 − AIC2 (2 state model). g PI(4,5)P2 dependency of the number of molecules adopting the slower mobility state (rectangle) and the longer binding state (circle) normalized to the total number of molecules. Ratio of the slower mobility state (D1, rectangle): P = 0.31 for 1 versus 5 mol% PI(4,5)P2; P = 0.13 for 1 versus 10 mol% PI(4,5)P2. Ratio of the longer binding state (k1, circle): P = 0.85 for 1 versus 5 mol% PI(4,5)P2; P = 0.02 for 1 versus 10 mol% PI(4,5)P2. P values were obtained by Welch’s t test. h Dissociation curve of single molecules on the membranes containing 1 mol% (magenta), 5 mol% (orange), or 10 mol% (blue) PI(4,5)P2. i PI(4,5)P2 dependency of the average dissociation rate constant (rectangle) and the dissociation rate constant for state 1 (circle) and state 2 (triangle). Welch’s t test; comparisons were made with “1 mol% PI(4,5)P2” column for the same state. Blue and red indicate D2 and average D, respectively. Data of 1, 5, and 10 mol% PI(4,5)P2 are the mean ± SD from 12 (4 experiments, 6000 molecules), 9 (3 experiments, 45,000 molecules), and 9 (3 experiments, 45,000 molecules) separate movies, respectively.
