Figure 1

validation of comparative RT-PCR. Comparison ofNorthern and RT-PCR analysis with H1-180 and H1-129. A shows Northern analysis of total RNA (5 μg) from control (C) or FSH-treated GCs (FSH). Hybridization was performed with H1-180 and H1-129 cDNA probes coming from forward hybridization (H1 = FSH-induced genes). The amount and integrity of RNA in each lane was checked with ethidium bromide staining of the gels before transfer (28 S lane). B shows the results of PCR amplification of H1-180 and H1-129 cDNAs using specific primers. Total RNA was extracted from control (C) or FSH-treated (FSH) cells and was reverse-transcribed. For PCR, four different amounts of each cDNA were used: a: 1 ng, b: 250 pg, c: 50 pg, d: 10 pg and 2 controls (-: water, +: corresponding insert). PCR amplification of I11a (plant external control added to each RNA before RT) was performed on the same samples (control and FSH-treated cDNAs) to check efficiency of the RT and PCR processes.