Quantstudio design analysis software version 1

Total RNA was extracted from cells using the Trizol® reagent (Invitrogen) and purified using the RNeasy Mini kit from Qiagen. 1 µg of purified RNA was used for cDNA synthesis using the High-Capacity cDNA Reverse Transcription Kit (ThermoFisher Scientific) according to the manufacturer's instructions. The QuantStudio 5 Real-Time PCR System (Thermo Fisher Scientific) was used to assess the expression levels of the mRNAs, miRNAs, and pseudogenes of interest. GoTaq® qPCR Master Mix (Promega) was used as a SYBR master mix reagent for the qPCR procedures. The qRT-PCR data was analyzed using the QuantStudioTM Design & Analysis Software Version 1.2 (ThermoFisher Scientific) and represented as relative expression(ΔΔCt), normalized against either GAPDH or β-actin. The primer sequences used for the quantitative real-time PCR are provided in Supplementary Table 1.

1 ug of RNA was reverse transcribed using Qscript cDNA Supermix (QuantaBio). The cDNAs were diluted 3 times for expression analysis. qRT-PCR on cDNA or ChIP-DNA were performed on 384 well plates on a QS5 system (Thermo Scientific) with GoTaq qPCR Master Mix (Promega, Madison, WI). The fold change or percentage input of the samples was calculated using the QuantStudioTM Design & Analysis Software Version 1.2 (ThermoFisher Scientific) and represented as relative expression (ΔΔCt). All measurements were performed in triplicate. Primers used in this study are listed in Supplementary Table S1.