RNA Isolation and qRT-PCR Analysis

The total RNA from BLs (n = 5 per group) was isolated with RNA isolation kit (PicoPure, Arcturus, ThermoFisher, Foster City, CA, USA) and used to synthesized cDNA with iScript reverse transcriptase (BioRad). The qRT-PCR analysis was performed as described by [38 (link)]. In a brief, the relative mRNA abundance of all genes was analyzed by real-time quantitative (q)RT-PCR with SYBR Green master mix using Cycler BioRad system. Threshold (Ct) values normalization of all tested genes was done with (Ct) values of GAPDH. The conditions used for PCR amplification were: initial denaturation at 94 °C for 5 min followed by 40 cycles of 94 °C for 30 s, 58 °C for 30 s, and 72 °C for 30 sec. For mRNA expression pattern analysis, three independent experiments were performed with four replicates. Primers used for RT-PCR and qRT-PCR are listed in Table S3.