Microarray suite version 5.0 mas 5.0

Yeast cells were grown in YPD medium to mid-log phase and were collected by centrifugation. Total RNA was extracted from whole cell lysates using hot phenol and RNA (200 ng) was used for first strand cDNA synthesis using the GeneChip^® 3′ IVT Express Kit (Affymetrix, Santa Clara, CA, USA). aRNA (7.5 μg) was hybridized for 16 hr at 45 °C in a GeneChip Yeast Genome 2.0 Array. GeneChips were washed and stained in a Affymetrix Fluidics Station 450, and then scanned using the GeneChip Scanner 3000 7G System. The data were analyzed using Microarray Suite version 5.0 (MAS 5.0) with Affymetrix default analysis settings.
The accession number of the microarray data is GSE220290.

Total RNA was extracted from FACS-sorted Treg using an RNeasy Mini Kit (Qiagen), according to the manufacturer’s instructions. The purity of the RNA was checked using a NanoDrop 2000 spectrophotometer (ThermoFisher Scientific) and a 2100 Bioanalyzer (Agilent Technologies). Biotinylated cRNA was synthesized from 250 ng of total RNA using the GeneChip 3′IVT PLUS Reagent Kit (Affymetrix), according to the manufacturer’s instructions. The yield of biotinylated cRNA was checked using the NanoDrop 2000 spectrophotometer. Following fragmentation, 15 μg of cRNA was hybridized to a GeneChip Mouse Genome 430 2.0 Array (Affymetrix) for 16 h at 45 °C. Arrays were washed and stained in a GeneChip Fluidics Station 450 (Affymetrix). Arrays were scanned using GeneChip Scanner 3000 7G. Single Array Analysis was calculated using Microarray Suite version 5.0 (MAS5.0), with the Affymetrix default setting and global scaling as the normalization method. The trimmed mean target intensity of each array was arbitrarily set to 500.