Fig. 1

Tat is palmitoylated in T-cells, macrophages and neuron precursors. a T-cells (Jurkat), macrophages (RAW 264.7) or neuron precursors (PC12 cells) were labeled overnight with 17-ODYA and Tat-His₆ in lipid-free medium before cell lysis, Tat-His₆ purification, biotin labeling of 17-ODYA using click chemistry and SDS-PAGE. The blot was first incubated with avidin-peroxidase to detect biotin, then with anti-Tat antibodies. b PC12 cells were transfected with Tat-FLAG before overnight labeling with 17-ODYA, anti-FLAG immunoprecipitation, and click chemistry. When indicated 100 µM 2-bromopalmitate (2-BP) was present during labeling with 17-ODYA. c PC12 cells were transfected with Tat-Flag before anti-Flag immunoprecipitation and acyl-biotin exchange. Hydroxylamine (HA) treatment is used to remove the acyl chain and enables to replace it with biotin for palmitoylation detection. Representative data from n = 3 experiments