RNA Extraction and qRT-PCR Analysis Protocol

RNA was extracted from BLs using an RNAqueous™-Micro Total RNA Isolation Kit (Ambion, Austin, TX, USA), according to the manufacturer’s instructions. The concentration of extracted total RNA was quantified by a NanoDrop 2000 Spectrophotometer (Thermo Fisher Scientific, Wilmington, DE, USA) and presented in Table S1. Using the RNA, complementary DNA (cDNA) was synthesized by a Maxime RT premix kit (iNtRON, Gyeonggi, Korea). qRT-PCR was carried out using a StepOnePlus Real-Time PCR System (Applied Biosystems, Foster City, CA, USA) and the protocol in detail was previously described [18 (link)]. The expression of target genes was measured and normalized relative to the control house-keeping gene, 18S rRNA [38 (link),39 (link),40 (link)]. The gene expression values were calculated as previously described [18 (link)]. The list of primers is presented in Table 1.