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Fig. 9 | BMC Biology

Fig. 9

From: Mechanisms of tethering and cargo transfer during epididymosome-sperm interactions

Fig. 9

Disruption of sperm lipid rafts compromises the efficacy of DNM1 translocation and epididymosome-sperm interaction. To examine the role of lipid rafts in mediation of epididymosome-sperm interactions, cells were pre-treated with mβCD to sequester membrane cholesterol and disrupt lipid raft integrity. Thereafter, spermatozoa were incubated with biotinylated (membrane impermeant) epididymosomes for 1 h. Spermatozoa were then fixed and subjected to immunofluorescence detection. a A significant reduction in the number of cells with post-acrosomal biotin labeling was observed in spermatozoa pre-treated with mβCD vs those of untreated controls. Post-acrosomal labeling was assessed in a minimum of 100 cells 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. **P < 0.01 compared to control. Individual data points for each replicate are provided in Additional file 7: Raw data. b, c Representative immunofluorescence images of triple stained caput spermatozoa: GM1 (red; lipid rafts), biotin (green), and DNM1 (blue). Compared to untreated control (b), mβCD treatment (c) elicited a loss of raft integrity with GM1 being heterogeneously dispersed throughout the sperm head. In these cells, DNM1 was mainly retained in the acrosomal domain, but did display a tendency to co-localize with GM1 and biotinylated proteins (white arrowhead)

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