IHC for XIAP
After incubation in TUNEL mixture, sections were immersed in rabbit polyclonal anti-human XIAP antibody (1:50) and, subsequently, in rho-damine-conjugated goat anti-rabbit IgG (1:200 in PBS, 1 h, RT). The XIAP signal (indicated by rhodamine) was generated with excitation and emission wavelengths of 568 and 630 nm, respectively. Confocal microscopic TUNEL and XIAP images were captured using NIH Image 1.61 software. generic Xenical online
All experiments were carried out three to four times. Following confirmation that no interreplicate differences existed in each experiment (oneway ANOVA), individual observations from all replicates were pooled for analysis by two-way (repeated-measure) ANOVA (PRISM software version 3.0; GraphPad, San Diego, CA). When XIAP content and the apo-ptotic DNA fragmentation were expressed as fold of control, they were arcsine square root-transformed before one-way or two-way ANOVA. Granulosa cell number was analyzed by two-way ANOVA. Differences between experimental groups were determined by the Tukey or Bonferroni posttest. The extent of granulosa cell apoptosis between experimental groups was analyzed by the chi-square test.