Fig. 6

Temporal gut-expression of selected hub and connecting transcripts in the 'blue' and 'turquoise' co-expression gene networks for non-exposed (NV) and tomato spotted wilt virus (TSWV) - infected (V) larvae of Frankliniella occidentalis. Droplet-digital PCR was performed to quantify normalized transcript abundance of A) FOCC013675 (lysosomal alpha-mannosidase, 'blue' hub), B) FOCC004976 (lysozyme, 'blue' hub), C) FOCC013756 (cytochrome p450: CYP3653A2, 'turquoise' hub), and D) FOCC001422 (putative beta-glucosidase, 'blue' connecting gene) during (DVE) and post virus exposure (PVE) at six time points during larval growth and development. Asterisks indicate statistically significant differences in gene expression level for pairwise comparisons between V and NV guts (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001). The corresponding abundance of E) TSWV nucleocapsid gene (N RNA) and F) TSWV non-structural small RNA gene (NSs RNA) in larval guts was also temporally monitored using real-time quantitative reverse-transcription PCR (qRT-PCR). Normalized abundance of each thrips and viral RNA was normalized to Frankliniella occidentalis actin RNA. Bars headed by different letters indicate statistically significant differences (P < 0.05) between time-points. No virus was detected in NV guts. Developmental stage (L1, L2) and time-stage corresponding to the RNA-Seq analysis of this study are delineated at the bottom of the figure; double dashes in the developmental stage timeline indicate the transition period (molting) from L1 to L2, thus the 30-h sample included both stages. Each expression data point/bar represents the mean and standard error of three biological replicates (independent experiments), and each replicate represents a pool of 60 larval guts