normalization data transformation protocol
Array Pro software was used to calculate differences in signal intensities and to normalize by Z-score transformation ID_REF = VALUE = Averaged z data
array scanning protocol
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RNA was denatured with NaOH and spotted onto S&S Nytran SuperCharge nylon membranes. Twenty-four 384-well plates were printed using a BioRobotics MicroGrid Arrayer.
RNA was reverse transcribed in the presence of [α-33P]dCTP.
nucleic acid extraction protocol
RNA was extracted with phenol/chloroform from C2C12 cell extracts that were subject to immunoprecipitation with an antibody that recognized either HuR or IgG.
sample treatment protocol
Proliferating C2C12 muscle cells were grown in DMEM supplemented with 20% FBS and 1% Pen/Strep. At 80% confluency, cells were collected.