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. 2017 Feb 6;114(8):E1365–E1374. doi: 10.1073/pnas.1612254114

Fig. 6.

Fig. 6.

Autoacylation of zDHHC enzymes by fatty acid azides. HEK293T cells transfected with HA-tagged zDHHC constructs were incubated with C14:0, C16:0, or C18:0 fatty acid azides for 4 h at 37 °C. Fatty acid azides were then labeled by click chemistry using an alkyne-800 infrared dye. Isolated proteins were resolved by SDS/PAGE and transferred to nitrocellulose membranes. Representative click signals and Western blots with positions of molecular mass standards indicated, together with quantified data (mean ± SEM) are shown for each zDHHC enzyme. (A) Autoacylation of zDHHC enzymes active against SNAP25B. zDHHC2, n = 6; zDHHC3, n = 14; zDHHC7, n = 11; zDHHC15, n = 6. (B) Autoacylation of additional zDHHC enzymes. n ≥ 6. ns, not significant; *P < 0.05; **P < 0.01; ***P < 0.001. The arrowhead on the zDHHC5 blot indicates the zDHHC5 band.