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Methoxychlor-Induced Atresia: MATERIALS AND METHODS(11)

A standard curve was generated from five serial dilutions of one of the samples, thus allowing analysis of the amount of cDNA in the exponential phase. FSHR primer sequences were: (forward) 5′-AGCAAGTTTGGCTGTTATGAG G-3′, (reverse) 5′-GTTCTGGATGATTTAGAGG-3′. An initial incubation of 95°C for 10 min was followed by denaturing at 94°C for 10 sec, annealing at 55°C for 10 sec, and a final extension at 72°C for 10 sec for 50 cycles followed by final extension at 72°C for 10 min. A standard curve was generated by the software, and p-actin was used for each sample as an internal standard. Final values for FSHR expression were calculated as the ratio FSHR: p-actin. Primers specific for the mouse p-actin were used as an internal control as previously described. All experiments were performed in triplicate.

Statistical Analysis

All data were analyzed using SPSS statistical software (SPSS, Inc., Chicago, IL). For all comparisons, statistical significance was assigned at P s 0.05. For comparisons between sesame oil-treated and MXC-treated mice and between genotypes, we used analysis of variance when comparing more than two groups and the Student f-test when comparing two groups. In the Western blot comparisons, we normalized doses to controls and conducted analysis of variance and linear regression analysis.