Activity-sensitive synaptic translocation of DHHC2. (A) No change in DHHC autopalmitoylation (detected by the ABE method) was seen upon activity blockade (TTX or Kyn) of hippocampal neurons, suggesting that DHHC activity remains constant. (B and C) TIRFM imaging revealed that treatment with Kyn or TTX translocated DHHC2-GFP near the plasma membrane. n = 3; *, P < 0.05; **, P < 0.01 compared with control. Kymographs (pseudocolor) represent the changes in the intensity of DHHC2-GFP over time. (D) The translocation of DHHC2-GFP induced by Kyn treatment was reversible upon washing out Kyn. (E and F) Colocalization of endogenous DHHC2 with PSD-95 steadily increased over prolonged TTX or Kyn treatment. (F) n = 5–7 each; **, P < 0.01; ***, P < 0.001. (C and F) Error bars indicate SD. Bars: (B) 2 µm; (D) 5 µm; (E [left]) 3 µm; (E [right]) 1 µm.