Extended Data Fig. 1: Detection of ‘high-single’ and ‘low-many’ var expression states in IT4.
(a) Clone tree of wildtype IT4 parasites. Each pie chart represents the var profile of an individual subcloned population determined by qRT-PCR, each slice of the pie represents the expression level of a single var gene. Expression of each gene is determined as relative to seryl-tRNA synthetase (PfIT_020011400). Populations are classified as ‘low-many’ if the expression level of no individual gene makes up more the 50% of the total var signal. The annotation number of the most highly expressed var gene is shown below the pie chart for populations expressing a dominant var gene. Vertical and horizontal lines delineate sequential rounds of subcloning by limiting dilution. (b) Total var expression levels as determined by qRT-PCR for all the subclones in (A). The mean ± SD interval is shown, and an unpaired two-tailed t-test indicates a ****p < 0.0001. (c) Relative expression of control genes as determined by quantitative real-time RT-PCR (qRT-PCR) and represented as relative to seryl-tRNA synthetase (PF3D7_0717700). The mean ± SD interval is shown, and an unpaired two-tailed t-test indicates non-significant differences (ns) between selected high-singles (n = 6) and low-many populations (n = 6). The control genes are: SBP1 (Skeleton Binding Protein 1, PfIT_050006700), MSP1 (Merozoite Surface Protein 1, PfIT_090034900), RFC (Replication Factor C, Subunit 1, PfIT_020024100), DNA Pol alpha (DNA Polymerase Alpha, Subunit A, PfIT_040016500).
