(A) Parasite growth curves in five volunteers infected by mosquito bite; each line represents one volunteer and the horizontal green line represents the lower limit of detection (five parasites ml−1). On days 9, 10, and 11 post-infection parasites were isolated from whole blood and immediately processed for ex vivo RNA-sequencing – for every volunteer, read counts were pooled from all three time-points to generate a comprehensive parasite transcriptome 2–3 cycles after liver egress. (B) Proportion of read counts that map to groups A, B, C, or E (var2csa) var genes after mosquito challenge (top row) or blood challenge (bottom two rows); parasite samples are colour-coded by host response. Note that the two inoculum samples are a technical replicate of a single biological sample. (C) Log2(cpm) expression values of var gene intron-spanning reads in the inoculum and diagnosis samples after blood challenge. Genes (columns) are ordered by hierarchical clustering and colour-coded by var type; parasite samples by host response.