RNA-seq after FACS enables the study of host responses to P falciparum at the terminal stages of erythroid differentiation. (A) Schematic diagram of experimental design for characterizing transcriptional host responses to P falciparum in specific erythroblast populations. Erythroblasts were removed from culture on day 7 or day 14 and mixed with GFP-expressing P falciparum schizonts or an equivalent volume of media. After 24 hours, infected and uninfected erythroblast populations were collected via FACS for RNA extraction and transcriptomic analysis by RNA-seq. (B-C) Principal component analysis of RNA-seq data set. Marker shape indicates erythroblast population. Color represents infection condition. (D) Heatmap showing Spearman correlation between the parasite transcriptome in nucleated erythroblasts and the intraerythrocytic parasite transcriptome measured every 8 hours, as published by Otto et al.42 Colored annotations represent cell type, and each column represents a single replicate. Hrs, hours.
