Cfx96 qpcr instrument

Total RNAs were isolated with QIAzol Lysis Reagent (Qiagen) and precipitated according to the manufacturer’s instructions. Reverse transcription was performed with 1 μg of total RNAs with the SuperScript^™ First-Strand Synthesis System for RT-PCR (Invitrogen). The resulting cDNA was used to perform gene expression analysis using the BioRad CFX96 qPCR instrument and Power SYBR Green PCR Master Mix (Thermo Fisher Scientific). The isoform-specific quantitative PCR (qPCR) primers are described in Supplementary Table S2.

Total RNAs were isolated from 661w cells with QIAzol Lysis Reagent (Qiagen) and precipitated according to the manufacturer's instructions. Reverse transcription was performed with 1 µg of total RNAs with the SuperScript First-Strand Synthesis System for RT-PCR (Invitrogen). The resulting cDNA was used to perform gene expression analysis using the BioRad CFX96 qPCR instrument and Power SYBR Green PCR Master Mix (Thermo Fisher Scientific). The following quantitative PCR (qPCR) primers were used to quantify OXR1 expression: OXR1_Fwd1: 5′-GCAAACTTTGGAAAACCCATACTA-3′, OXR1_Rev1: 5′-GATGCCTTTGCTTTTCTTTTAAAAG-3′, OXR1_Fwd2: 5′-CAAACTTTGGAAAACCCATAC-3′, OXR1_Rev2: 5′-AACTTATGTAATCGATGCC-3′, β-Actin_Fwd: 5′-GGCTGTATTCCCCTCCATCG-3′, and β-Actin_Rev: 5′-CCAGTTGGTAACAATGCCATG-3′.