Fig. 10

Pharmacological inhibition of dynamin does not compromise lipid raft or DNM1 translocation. To assess the impact of dynamin inhibition on lipid raft and DNM1 localization, caput spermatozoa were pre-treated with either the dynamin inhibitor, Dynasore, or the DMSO vehicle control. Thereafter, spermatozoa were incubated with biotinylated (membrane impermeant) epididymosomes for 1 h. Spermatozoa were then fixed and subjected to immunofluorescence detection. a, b Representative immunofluorescence images of triple stained caput spermatozoa are presented from DMSO and Dynasore-treated groups: G_M1 (red; lipid rafts), biotin (green), and DNM1 (blue). However, due to the difficulty of counting the triple-stained cells, a subset of spermatozoa from each of the DMSO and Dynasore treatment groups were individually labeled for either c G_M1 (red; lipid rafts) or d DNM1 prior to assessment of post-acrosome labeling. A minimum of 100 cells were assessed per treatment group, with these experiments being replicated three times. Each replicate comprised pooled material from at least three mice. The results are presented as the mean ± S.E.M. n.s. not significant compared to DMSO control. Individual data points for each replicate are provided in Additional file 7: Raw data