FIG 1.

Coinfection workflow for mass spectrometry-based proteomics profiling. BALB/c macrophages were cocultured with C. neoformans H99 (Cn) for 90 min followed by removal of nonphagocytosed cells (i.e., immediate C. neoformans samples) (time point 1). After 48 h, established C. neoformans samples were collected (time point 2), and designated C. neoformans-infected macrophages were cocultured with K. pneumoniae (Kp) for 90 min followed by sample collection (i.e., established C. neoformans plus K. pneumoniae immediate) (time point 3). After 24 h, established C. neoformans plus K. pneumoniae established samples were collected (time point 4). Additionally, K. pneumoniae-only-infected macrophages were collected after immediate (90 min) (time point 5) and established (time point 6) infections. All samples were subjected to our total protein extraction protocol, followed by digestion, measurement on the mass spectrometer, and data processing and analysis using MaxQuant and Perseus. The experiment was performed in biological quadruplicate. The figure generated with Biorender.com.