RT-qPCR
Cells were grown for 48 hours after lipofection before being harvested using RNAprotect (Qiagen). Total RNA was extracted using Direct-zol RNA microprep kit and quantified. Each condition was performed in triplicate using 50 ng of total RNA. For each 20 ul reaction, RNA was added to Luna Universal One-Step RT-qPCR master mix (New England Bio Labs) and two RT-qPCR primers at a final concentration of 0.4 µM. The following amplification cycle was run on a QuantStudio 3 real-time PCR machine (ThermoFisher Scientific): 55 °C for 10 min, 95 °C for 1 min, followed by 95 °C for 10 sec and 60 °C for 1 min for 40 cycles. All genes were normalized to beta Actin (ACTB) and expression was determined via relative quantification to lipofectamine only carrier control. Gene specific primers for each gene of interest were ordered from Integrated DNA Technologies and are as follows: ACTB: 5’-CACCATTGGCAATGAGCGGTTC-3’ and 5’-AGGTCTTTGCGGATGTCCACGT-3’, TBK1: 5’-GGATCACTGCCATTTAGACCC-3’ and 5’-CAGGCATGTCTCCACTCCAG-3’ (PrimerBank 309747068c3), KRT14: 5’- TGAGCCGCATTCTGAACGAG-3’ and 5’-GATGACTGCGATCCAGAGGA-3’ (PrimerBank 83641893c1), HNRNPA1: 5’-TCAGAGTCTCCTAAAGAGCCC-3’ and 5-ACCTTGTGTGGCCTTGCAT-3’ (PrimerBank 83641893c1).