Continuous palmitate cycling on PSD-95 nanodomains is mediated by local palmitoylating activity. (A) Dendritic regions of transfected hippocampal neurons were bleached, and then the fluorescent recovery of PSD-95-GFP or PF11-GFP at the individual clusters was monitored. Right, representative magnified images before bleaching and at 60 min after bleaching. Note that the recovered PF11-GFP kept its initially distinguished nanodomain locations (arrowheads). Asterisks, diffuse fluorescent signals of PF11-GFP in dendritic shaft that were subtracted from the FRAP at the clusters. Bars: (left) 5 µm; (right) 0.5 µm. (B) Sample FRAP of PF11-GFP at subsynaptic clusters. (C) Average FRAP of PF11-GFP (red), PF11-GFP treated with 2-BP (green), and PSD-95-GFP (blue). n = 3 experiments (5–6 clusters/experiment). (D) Fluorescence recovery of PSD-95-GFP and PF11-GFP at 60 min after bleaching in the presence or absence of 2-BP. ***, P < 0.001 by one-way ANOVA with post-hoc Tukey’s test (n = 3 experiments [5–6 clusters/experiment]).