Transcriptional changes in J774.16 macrophage like-cells to fungal infections, C. gattii-infected, replicate 1
J774.16 macrophage-like cells were seeded in culture plates in a density of 106 cells per milliliter and activated with 100 U/ml recombinant murine IFN- (Sigma Aldrich) and 500 ng/mL LPS (Sigma Aldrich) overnight.  Cells were washed with PBS and incubated for further 6 h in DMEM supplemented with 10% heat-inactivated fetal bovine serum in the presence of 107 Cryptococcus gattii R265 cells opsonized with the anti-GXM antibody 18-b7. The macrophages were lysed with TRIzol. The macrophage RNA was purified using the RNeasy Mini kit and the poly (A) RNA were purified from macrophage cells with Dynabeads mRNA purification kit. The RNA quality was assessed with Bioanalyzer 2100 system and sequenced in an Ion PGM System.