RT-PCR

RNA was isolated with a PureLink RNA Minikit (Thermo Fisher Scientific). cDNA was transcribed with the iScript Advanced cDNA Synthesis Kit (Bio-Rad). Preamplification was performed for 10 cycles with the SSO Advanced PreAmp Supermix (Bio-Rad). ARG2 was measured with primers 5′-GAAGAAATCCGTCCACTCCGT-3′, 5′-TTAGGTGGCAGCCCAAACT-3′; ARG1 was measured with primers 5′-TGGACCCTGGGGAACACTAC-3′, 5′-CGAGCAAGTCCGAAACAAGC-3′, developed by NCBI Primer-BLAST (70); and EIF3L was measured with primers 5′-TGACCCCTACGCTTATCCCAG-3′, 5′-GTTTGCTGTTCATACTGACGTTC-3′ (PrimerBank ID 339275830c1) (71), with SsoAdvanced Universal SYBR Green master mix (Bio-Rad) on a StepOnePlus PCR System (Applied Biosystems).