Fig. 7

Analysis of the involvement of DNM2 in epididymosome-sperm interaction. a Spermatozoa were incubated with biotin (membrane impermeant)-labeled epididymosomes for 1 h before being subjected to immunofluorescence detection of biotin (green) and DNM2 (red). Representative immunofluorescence images of the different sperm labeling patterns detected after this period of co-incubation are provided to illustrate the labeling of DNM2 in the acrosomal domain and minimal co-localization with transferred biotinylated proteins. Indeed, only relatively weak DNM2 labeling was detected in the post-acrosomal domain of those cells that incorporated abundant biotinylated proteins. b The relative abundance of DNM2 was quantified by immunoblotting of sperm homogenates in naïve cells (Sperm only) as well as those exposed to co-culture with epididymosomes (Sperm + ES). For the purpose of comparing the relative abundance of DNM2, band intensity was normalized relative to that of α-tubulin, with sperm only control nominally set to a value of 1 (n = 3). Individual data points for each replicate are provided in Additional file 7: Raw data. These experiments were replicated three different times with each sample representing pooled material obtained from at least three mice