*2.5. RNA Isolation and Reverse Transcription*

Cells were lysed in TRI reagent (Invitrogen, 11312940, Carlsbad, CA, USA). After this, 10% bromochloropropane (Sigma-Aldrich, B9673, Saint Louis, MO, USA) was added. Lysates were inverted and incubated at room temperature for 5 min and centrifuged (4 ◦C, 12,000 g, 10 min). The RNA containing supernatant was transferred to a new centrifuge tube and washed with 100% ethanol 2:1. RNA was further isolated using the RiboPure RNA purification kit (Thermo Fisher Scientific, AM1924, Waltham, MA, USA). Then, 500 ng RNA and 4 μL SuperScript VILO Mastermix (Invitrogen, 12023679, Carlsbad, CA, USA) were diluted to 20 μL in RNAse free water and reverse transcription was performed in a 2720 thermal cycler (Applied Biosystems, Foster City, CA, USA), using the following program: 10 min 25 ◦C, 60 min 42 ◦C, 5 min 85 ◦C. The cDNA was diluted 10x in RNAse free water.
