Sorted GFP+ BV2 cells after overexpression or knock-down of PU.1 were collected as described for western blotting. Cell pellets were lysed in QIAzol reagent and RNA was isolated with RNAeasy Mini kit according to the manufacturer’s instructions (Qiagen) including the Dnase treatment step with RNase-free DNase set (Qiagen). Quantities of RNA were measured using Nanodrop 8000 (Thermo Scientific) and reverse transcription was performed with 1–2 μg of total RNA using High-Capacity RNA-to-cDNA kit (Thermo Fisher Scientific). qPCR was performed on QuantStudio 7 Flex Real-Time PCR System (Thermo Fisher Scientific) using Power SYBR Green Master Mix (Applied Biosystems) with one-step PCR protocol. 3 ng of cDNA was used for all genes except Ms4a4a when 24 ng of cDNA was used in a 10 μl reaction volume. Primers were from PrimerBank80 or designed using Primer-BLAST program (NCBI) and are listed in Supplementary Table 14. Ct values were averaged from two technical replicates for each gene. Geometric mean of average Ct for the housekeeping genes GAPDH, B2M and ACTB was used as a reference that was subtracted from the average Ct for a
