Fig. 3

Time Course of Protein Palmitoylation Levels in Sperm. (A) Protein palmitoylation levels at different time points. Palmitoylation of total proteins extracted from sperm was evaluated using the ABE method with different durations of HTF incubation. The loading controls were confirmed by Western blotting. Bands in lanes with NH₂OH represent palmitoylated proteins. (B) Quantification of the level of protein palmitoylation, with band gray intensities indicated by arrows, was analyzed using the ImageJ software and normalized to the total amount of loaded protein. Data are presented as means ± SEM (n = 3, three repeated measures from different samples). A P-value < 0.05 denotes statistical significance. (C) Schematic diagram of the ABE method. NEM, N-ethylmaleimide; NH₂OH, Hydroxylamine; BMCC-Biotin, 1-Biotinamido-4-[4’-(maleimidomethyl)cyclohexanecarboxamido] butane