Research Paper Volume 11, Issue 10 pp 2968—2997

Figure 7. Expression of HSET protein in young and aged oocytes. Immunofluorescence analysis of HSET in young and aged GV, MI, and MII stage oocytes was utilized to determine whether an age-related decrease in Kifc1 and Kifc5b transcript abundance presages an equivalent decrease in protein abundance. Oocytes were labelled with anti-HSET antibodies followed by goat anti-rabbit 633 Alexa Fluor-conjugated (grey) secondary antibodies. Oocytes were then counterstained with the nuclear stain Hoechst 33342 (blue) and viewed using confocal microscopy. Scale bar = 20 μm. These experiments were repeated using three independent biological replicates, with each comprising a minimum of 10 oocytes, and representative images are shown. The immunofluorescence intensity of the entire cell was calculated for each oocyte as described in the Materials and Methods, and the mean of each biological replicate values ± SEM are shown. Statistical analyses were performed using Student’s t-test, * p < 0.05, ** p < 0.01. AU, arbitrary units.