2.6. Quantitative PCR (human AT biopsies)

qPCR analyses were performed as previously described [25]. TRIzol (Gibco BRL/Life Technologies, Roskilde, Denmark) were used to extract RNA. The PCR reactions were performed in duplicate using a LightCycler SYBR Green master mix (Roche Applied Science, Penzberg, Germany). β2 microglobulin was used as the housekeeping gene. Messenger RNA levels of β2 microglobulin were similar in both interventions. The fold change was calculated using the 2^–ΔΔCt method; ΔΔCt is the difference between the average Ct for the target gene and the house keeping gene at time x minus the difference between Ct for the target gene and the house keeping gene at baseline (GH bolus study) or at the control day (GH infusion study). Detailed information regarding primer sequences is shown in the supplementary data.