Comprehensive RNA Extraction and qPCR Analysis

Total RNA was extracted using TRIzol reagent (catalog number: 15596026, Invitrogen, Thermo Fisher Scientific, Carlsbad, CA, USA) and cDNA synthesis was performed using 1 ug RNA and High-Capacity RNA-to-cDNA kit (catalog number: 4388950, Applied Biosystems™, Thermo Fisher Scientific, Carlsbad, CA), according to the manufacturer’s instructions. Real-time PCR was performed using PowerUp™ SYBR™ Green Master Mix (catalog number: A25743, Applied Biosystems ™, Thermo Fisher Scientific, Carlsbad, CA, USA) on QuantStudio™ systems with respective software (Applied Biosystems™, Thermo Fisher Scientific, Carlsbad, CA, USA). Gene expression was normalized with CypA, Rps9, Gak, and Srp72 as housekeeping genes by using geNorm application [41 (link)]. Primer sequences are described in Table S1.