<p>(A–B) RT-PCR analysis of RNA extracted from the pituitary and other tissues from mice of the six different TG lines. <i>Gapdh</i> mRNA was used as a control. Ht, heart; Li, liver; Sp, spleen; Lu, lung; Ki, kidney; Br, brain; Pi, pituitary; Ov, ovary; Ut, uterus; Mu, muscle; In, intestine; M, 100 bp DNA ladder; P, upper and middle lanes, expression vector as a positive control and, lower lane, pituitary of a WT mouse; N, upper and middle lanes, pituitary of a WT mouse as a negative control and, lower lane, double-distilled water. (C) Northern blot analysis of transgene expression in adult tissues. Northern hybridization with the indicated cDNA probes was carried out using total RNA (20 µg) isolated from different tissues of TG and WT mice. Ethidium bromide (EtBr) staining of 28S, 18S, and 5S rRNA served as a control for RNA quality. Pituitary samples from four TG animals were detected. (D–E) <i>pFSHα</i> and <i>pFSHβ</i> mRNA levels in the pituitaries of TG and WT mice were analyzed using quantitative real-time PCR with specific primers and are expressed relative to β-actin (internal control). Data were combined from three independent experiments; bars represent means ± SEM (<i>n</i> = 7–8 mice per group).</p
