Adar2

Gene-specific exon-exon boundary PCR products (TaqMan gene expression assays, Applied Biosystems) were measured by means of a PE Applied Biosystems PRISM 7700 sequence detection system during 40 cycles. β-actin mRNA was used for normalization and relative quantification of gene expression was performed according to the 2-ΔΔCt method. Expression levels were represented in arbitrary units calculated as a relative-fold increase compared to the control sample arbitrarily set to 1. Quantitative RT-PCRs were repeated in triplicates from at least two independent experiments.
The primers were supplied by Applied Biosystems: OPHN1, ID Hs00609994_m1; ADAR1, ID Hs00241666_m1; ADAR2, ID Hs00953730_m1; β-actin, ID Hs99999903_m1. All the qRT-PCR data was also confirmed using the SYBR green method (Invitrogen) (data not shown).

Here, 1 μg of total RNA (pre-treated with DNase I) was used to generate cDNA by the ImProm-II Reverse Transcription System (Promega Corporation, Madison, WI, USA) using random hexamer primers according to the manufacturer’s instructions. GAPDH and/or β-Actin were used as controls for the normalization of mRNAs. The relative amount of each substrate was calculated by the 2^−ΔΔCt method [44 (link)]. Expression levels were represented as a relative-fold increase compared with the control sample, arbitrarily set to 1. All qRT-PCR reactions were performed in duplicates and repeated at least two times from independent experiments. All of the primers were supplied by Applied Biosystems: ADAR2, ID Hs00953730_m1; GAPDH, ID Hs99999905_m1; β-actin, ID Hs99999903_m1; ADAM12, ID Hs01106101_m1; and PTX3, ID Hs00173615_m1.