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Fig. 5 | BMC Biology

Fig. 5

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

Fig. 5

DNM inhibition reduces the transfer of epididymosome protein cargo to spermatozoa in vitro. a Spermatozoa were pre-treated with DMSO (vehicle control) or an equivalent concentration (100 μM) of Dynasore or Dyngo-Ɵ (an inactive isoform control) for 30 min, before being subjected to incubation with biotin (membrane permeant)-labeled epididymosomes for 3 h. In this assay, the ratio of epididymosomes to sperm was adjusted such that epididymosomes equating to a single mouse were co-incubated with pooled spermatozoa from four mice. Cell lysates were then subjected to Western blotting to detect the efficacy of biotinylated protein cargo transfer. b The pixel intensity of biotinylated protein bands detected within each lane were quantified using ImageJ software, and a representative trace of this analysis is included alongside the blot. a After imaging, blots were stripped and re-probed with anti-α-tubulin antibodies to confirm equivalent protein loading. These experiments were replicated three times with each replicate containing pooled sperm lysates from three mice, and a representative blot is presented

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