Elucidating the Role of the Desmosome Protein p53 Apoptosis Effector Related to PMP-22 in Growth Hormone Tumors

An Affymetrix Human Gene 1.1 ST array plate (Thermo Fisher Scientific) was hybridized with 2.3 µg of fragmented and labeled single-stranded complementary DNA (cDNA) Our cohort consisted of 21 GH tumors (11 DG and 10 SG) and 20 samples of normal human pituitary. Total RNA was isolated using TRIzol method (Invitrogen, Carlsbad, CA), followed by clean-up with a QIAGEN Bioinformatics RNeasy mini kit (Redwood City, CA). RNA was quantified by spectrophotometry, and RNA integrity was verified using the Agilent 2100 bioanalyzer (Agilent Technologies, Santa Clara, CA). Microarray targets were prepared and labeled from 150 ng of total RNA using the Ambion WT (Thermo Fisher Scientific, Waltham, MA) expression kit following the manufacturer’s instructions. An Affymetrix Human Gene 1.1 ST array plate (Thermo Fisher Scientific) was hybridized with 2.3 µg of fragmented and labeled single-stranded complementary DNA (cDNA). Hybridization and scanning were done on the Affymetrix GeneTitan instrument. Hybridization intensities were quantified and normalized across all arrays using the robust multichip average algorithm available as an array processing tool on Partek Genomics Suite software (Partek, St. Louis, MO).