Fig. 3.

Overexpression of CYP11A1 and NUSAP1 and their encoded proteins in human gonadotroph tumors. a RNA was extracted from frozen pituitary tumors obtained after transsphenoidal surgery. qRT-PCR was performed using TaqMan primer and probe sets specific to human NR5A1, CYP11A1, NUSAP1 and NEUROD1. The relative mRNA expression level of the target genes was normalized for input RNA using human TBP gene expression (housekeeping gene) and a calibrator human brain RNA always run in parallel and was calculated with the 2^−ΔΔCt formula. The obtained relative value was normalized against the average expression of normal pituitary arbitrarily set to 1. The boundary of the box closest to zero indicates the 25th percentile, the line within the box marks the median, and the boundary of the box farthest from zero indicates the 75th percentile. Error bars above and below the box indicate the 99th and 1th percentiles. b Human normal pituitary (top) and gonadotroph adenoma samples were used. IHC was performed using antibodies against P450scc (left) or against NuSAP (right) and counterstained with hematoxylin. Original magnification: ×200; insets: ×400. c Scoring of the expression of NuSAP, Ki-67 and P450scc in a series of human gonadotroph adenomas