Fig. 3.

Comparative analysis of mouse epididymosome protein abundance. The differential accumulation of epididymosome protein cargo was assessed via cyanine-dye labeling of epididymosome protein extracts, recovered from (A,) the proximal (caput versus, corpus) and (B,) distal (corpus versus, cauda) epididymal segments. Labeled proteins extracted from the two different epididymosome populations were mixed, prepared for resolution by 2D SDS-PAGE and visualized via the use of a Typhoon FLA 9500 laser scanner. This experiment was replicated three times using paired epididymosome samples (i.e., either caput and corpus or corpus and cauda epididymosomes) and depicted are representative merged gel images for (A,) caput (cyanine3-labeled, pseudo-colored in green) and corpus (cyanine5-labled, red) epididymosome proteins or alternatively, (B,) corpus (cyanine3-labeled, pseudo-colored in green) and cauda (cyanine5-labled, red) epididymosome proteins. C–E,, The relative abundance of epididymosome proteins was also determined via assessment of TMT reporter ion intensity in samples isolated from the caput, corpus and cauda segments of the mouse epididymis. For this analysis a threshold of ± 1.5 fold change (p, < 0.05) was set as the basis for assignment of differentially accumulated proteins. C,, The overall number of proteins that experienced no-change (black columns), increased (red columns), or decreased (green columns) accumulation in each epididymosomes population are shown. Similarly, the conservation of proteins that were either (D,) unchanged or (E,) experienced ≥ 1.5-fold increase (red font, ↑) or decrease (green font, ↓) change among different epididymosome sub-populations are also shown.