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. 2016 Jun 15;36(24):6431–6444. doi: 10.1523/JNEUROSCI.0419-16.2016

Figure 1.

Figure 1.

Screening for PSD-95 depalmitoylating enzymes. A, B, Isolated serine hydrolase (SH) clones were individually cotransfected with PSD-95-GFP into HEK293T cells (A) and COS7 cells (B). After metabolic labeling with [3H]palmitate, proteins were separated by SDS-PAGE, followed by fluorography (top) and Western blotting (WB) with GFP antibody (bottom). Arrowheads indicate the position of PSD-95-GFP. WT, Wild-type; C3,5S, mutated at cysteines 3 and 5 to serines. Normalized palmitoylation levels of PSD-95 relative to control (%) in COS7 cells are indicated (B). C, Effects of SH clones on palmitoylation levels of PSD-95 in HEK293T cells. PSD-95-GFP palmitoylation level was normalized by the expression level of PSD-95-GFP. Note that ABHD1, 3, and 4 reduced the PSD-95-GFP expression. n = 3 independent experiments. D, Expression of individual FLAG-tagged SH clones was confirmed. Asterisks indicate bands detected by overexposing the film.