Fig. 1: Half of the salivary gland transcriptome is rhythmic.

a, The experimental design from two independent experiments for the sequencing of salivary glands transcripts from A. stephensi mosquitos infected with P. berghei. Infected mosquitos were maintained in cyclic conditions (12 h light/12 h dark). Eighteen days after mosquito infection, mosquitos were segregated into light/dark (LD) and constant dark (DD) conditions. Twenty-four hours later, mosquitos were dissected every 4 h over 3 days (72 h) (n = 32 samples for each light/dark (LD) and dark/dark (DD) condition). b, Representation of three genes from the top 20 cycling genes (based on significance) from each condition and the circadian-algorithm-fitted lines. c,d Hierarchical clustering and heat maps for light/dark (c) and dark/dark (d) dataset by reordering of the timepoints according to similarity in gene expression. e, Heat map of cycling genes from salivary glands from mosquitos in light/dark and dark/dark. Each row represents a gene. Gene expression is z-scored. f, The distribution of the peak of expression for all cycling genes in LD and DD conditions. This shows two main times of day where most of the cyclic transcription occurs. Most gene expression peaks at 4 h after lights turn on, or the beginning of daytime; and 4 h after lights off, meaning 16 h after the beginning of daytime. g, The phase (time of peak of expression) for each of the 2,360 common genes is largely maintained across conditions (LD and DD). The plot represents a double plot of the phase of each cycling gene in each condition. Red dashed lines denote linear correlation between the phases of 2,360 common cycling genes in each condition. Solid grey lines denote 0, 24 or 48 h timepoints on each axis. h, The circadian fold change in expression of the common cycling genes across conditions. The violin plot width depicts distributions of fold-change values. ks, Kolmogorov–Smirnov statistical test. Mosquito icon in panel a designed by Freepik.

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