Fig. 2: Salivary-gland-specific transcripts cycle throughout the day.

a, The rhythmic expression profile of nine well-characterized salivary gland proteins known to have a role in bloodmeal efficiency and/or malaria transmission (Saglin, TRIO and GILT). b, The expression profile of 21 transcripts encoding for proteins associated with blood feeding, identified by being differentially expressed between female and male salivary glands⁹⁴. Nineteen of these genes show a similar phase and profile of gene expression. Refer to Extended Data Fig. 8. Grey bars in the left plot represent nighttime or 12 h of lights off. c, The top significant biological functions cycling under both LD and DD conditions at specific times of day. Representative cycling genes from the pathway are from the DD condition. d, Representation of the glycolysis and gluconeogenesis pathway with the genes that code for the enzymes inside rectangular boxes. Only 2 out of the 11 represented genes of the glycolysis pathway do not cycle in the mosquito salivary glands throughout the day (unfilled boxes), whereas the other 9 cycle (coloured boxes). e,f, Normalized expression profile (fold-change to mean RPKM expression values across timepoints) for each gene from the DD condition belonging to the glycolysis and gluconeogenesis pathway that cycle with a maximum expression between 0 h and 4 h (e), and from pentose and glucuronate interconversions (inter.) between 4 h and 8 h (f). g,h, Normalized protein expression levels (log₂ intensities − log₂ median) from two salivary gland proteins across four timepoints. Normalized protein expression levels (log₂ intensities − log₂ median) were computed by subtracting the log₂ median intensity value of that protein from each of its log₂ intensity values from each replicate in each timepoint. Each point represents the normalized intensity of that protein from each replicate—a pool of five salivary glands, with three replicates per timepoint. Differential expression analysis in Extended Data Figs. 3 and 4 and Supplementary Table 1f with significance testing and fold-change calculation using MSstats. One-way analysis of variance (ANOVA), one-sided, **P = 0.0016 for ASTE008071 (g) and **P = 0.0096 for ASTE004394 (h).

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