After 48 h, H99S Cn cells were collected from the RPMI media, centrifuged, and stored in RNA Later at –80 °C overnight. The next day, the RNA Later was removed and total RNA was extracted using an RNeasy Mini Kit (Qiagen, Hilden, Germany) after homogenization of Cn cells in a Mini Beadbeater 16 (Biospec Products, Bartlesville, OK, USA). After 48 h of incubation, male or female macrophages were collected and lysed using a cell scraper and then any remaining cells were lysed using an 18G needle 10×. Total RNA was then isolated using an RNeasy Mini Kit (Qiagen, Hilden, Germany). For all samples, genomic DNA contamination was removed from the total RNA using a Message Clean Kit (GeneHunter, Nashville, TN, United States), and the RNA concentration was measured using a NanoDrop 2000 Spectrophotometer (Thermo-Scientific, Waltham, MA, United States). RNA quality was determined using a Bioanalyzer (Agilent Technologies, Santa Clara, CA, USA) and samples with a RIN value >8 were sent to Novogene Corporation (Sacramento, CA, USA) for RNA sequencing. A total of 100 ng of total RNA was used for each sample, per Novogene's instructions. Done at Novogene Corporation for all samples.