Figure 4.

P2X7R mutants show altered diffusion. K17A and K64A mutants were generated by sight-directed mutagenesis from the wild-type (WT) P2X7-Dendra2 receptor plasmid. sptPALM was performed and diffusion analyzed. (A) Compared to WT-P2X7Rs, K17A-mutant showed slower diffusion, while K64A-mutant showed faster diffusion for total (100%), “free” (~65%) and “trapped” (~35%) population. Plotted data shows a distribution from 25 to 75th percentile and the median value (Kolmogorov-Smirnov statistical test ^***p < 0.001, ^**p < 0.01). (B) Median diffusion coefficient and number of trajectories for each population. (C,D,E) MSD plot shows an increased confined diffusion in K17A-mutant receptors and a decreased confined diffusion in K64A-mutant receptors with respect to wild-type P2X7Rs.