Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - (SupplemFig1.tif; 1.0 MB) Immunocytochemical localization of ceramide in cumulus-enclosed oocytes (COC) from ASMase WT and KO female mice. COC from young WT females (A-C) exhibited high levels of ceramide in contrast to COC from young KO females (D-F) which showed extremely low basal levels of ceramide prior to culture. Pixel intensity in oocytes from ASMase WT was 100% (n=25) compared to 65 ± 4.64% decrease in PI for oocytes from ASMase KO (n=30). The apparent discrepancy in size between oocytes from ASMase WT and KO is due to an artifact in slide preparation; different pressures were inadvertently applied to the coverslips, but it doesn't affect the results.
• Supplemental Figure 2 - (SupplemFig2.tif; 285 KB) Percent apoptosis in cumulus-enclosed oocytes (COC) of aged mice in the presence of a Bcl-2 homology domain-4 (BH4) peptide. COC obtained by superovulation of aged female ICR mice were cultured in vitro for 24 h, without or with BH4 (15 μg/ml) after which the percentage of oocytes (of the total number cultured in each group) exhibiting morphological evidence of apoptosis was determined. Treatment of COC from aged mice with the BH4 peptide suppressed the age-related increased in the incidence of oocyte apoptosis. The data presented represent the mean ± SEM of results from four independent experiments, with each incorporating at least three mice per group. The total number of oocytes analyzed per group is provided above the respective bar. *, P<0.05, compared with respective control value.