qPCR
U2OS cells were seeded on 10 cm plates under standard culture conditions and incubated for 24-48 hours to reach 100% confluency. Once cells reached confluency, RNA from cells were isolated using RNeasy Plus Mini Kit (74134, Qiagen). Using the RNA, cDNA was synthesized using qScript cDNA SuperMix and protocol from Quantabio (101414-102). The qPCR primer cocktails were made using PerfeCTa SYBR Green FastMix (101414-276), forward primer, reverse primer (Real Time Primers) and RNase/DNase free H2O. In a 96 well qPCR plate, 18 µl of primer cocktail was added and 2 µl of cDNA, accordingly. qPCR was run using BioRad cfx96 and fold gene expression was calculated.