RNA was isolated from cells using TRIzol (Thermo Fisher Scientific) according to the manufacturer’s instructions. One microgram of total RNA was used for complementary DNA (cDNA) synthesis. For standard mRNA analysis, oligo(dT)20 (Invitrogen) was added. For splice variant analysis, RNA was deoxyribonuclease (DNase)–treated (Sigma-Aldrich) before cDNA synthesis using random hexamers (Invitrogen). Moloney Murine Leukemia virus (M-MLV) reverse transcriptase (Promega) was used as per the manufacturer’s instructions. Quantitative reverse transcription PCR (qRT-PCR) was carried out in triplicate using the indicated primer pairs and the Brilliant III SYBR Green qPCR Master Mix (Stratagene) on an MX3005P (Agilent) qPCR instrument. Results were expressed as average (mean) fold change compared to control treatments using the ΔΔCt method from three biological repeat samples. Glyceraldehyde-phosphate dehydrogenase or actin primer sets were used as an internal calibrator. Error bars represent SE unless otherwise indicated.

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