Fig. 2

NPY mRNA abundance and protein content of granulosa cells is higher in EA than LAF. Granulosa cells of PA, EA and LAF were isolated separately using rats primed with eCG, as described in the methods section. a, c Total protein was extracted and NPY contents were assessed by Western blot. Representative Western blot result is shown a. NPY protein content of granulosa cells in PA and EA was higher than that in LAF c. b Total mRNA was extracted, reverse transcribed and amplified by real-time PCR using specific primers for Npy and Gapdh. Expression of Npy mRNA was normalized to RNA loading for each sample using Gapdh mRNA as an internal standard. Npy mRNA of granulosa cells in EA was higher than that in LAF. These results suggest that granulosa cell NPY is more important for pre-antral and early-antral follicles development than at later stages. Each granulosa cell replicate was isolated from at least two rats. Results are expressed as mean ± SEM of independent replicates (A, n = 4; B, n = 5) and analyzed by one-way ANOVA and Tukey’s multiple comparison. *, p < 0.05 (vs. EA); †, p < 0.05 (vs. PA)