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Fig 1.

Two-step bioassay to determine the effect of the suppression of the endocytic genes on the expression of laccase2 in WCR adults.

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Fig 2.

Relative transcript levels of the endocytosis-related genes after dsRNA exposure.

Relative transcript levels of Chc (A and B), Vha16 (C and D), AP50 (E and F), Arf72A (G and H), and Rab7 (I and J) genes in WCR adults after the first (stage 1) and second (stage 2) dsRNA exposure evaluated by RT-qPCR. Values shown are the means and standard errors (±SE) of three biological replicates each with two technical replicates. Different letters represent significant differences at p-value < 0.05.

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Fig 3.

Effect of the knockdown of the endocytosis-related genes on the RNAi response.

Effect of the knockdown of the AP50 (A), Chc (B), Vha16 (C), Rab7 (D), and Arf72A (E) on the relative transcript level of laccase2 was evaluated by RT-qPCR analysis. Values shown are the means and standard errors (±SE) of three biological replicates each with two technical replicates. Different letters represent significant differences at p-value < 0.05.

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Fig 4.

Two-step bioassay to determine the effect of the suppression of silA and silC genes on the mortality and V-ATPase A expression in WCR adults.

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Fig 5.

Relative transcript level of silA, silC and laccase2 genes after dsRNA exposure.

Relative transcript level of silA (A), silC (B) and laccase2 (C) evaluated seven days after the injection of 600 ng of the respective dsRNA or a combination of silA and silC dsRNAs into WCR adults. Values shown are the means and standard errors (±SE) of three biological replicates, each with two technical replicates. Different letters represent significant differences at p-value < 0.05.

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Fig 6.

Mortality of WCR adults and relative V-ATPase A expression.

(A) Mortality of WCR adults from the different treatment groups after 14 days. WCR adults were injected with 600 ng of GFP, laccase2, silA, silC, or a combination of silA and silC dsRNAs and subsequently fed with V-ATPase A dsRNA. WCR adults injected with GFP and not exposed to V-ATPase A dsRNA were used as controls. (B) Relative V-ATPase A transcript levels were evaluated by RT-qPCR seven days after exposed to V-ATPase A dsRNA. Values shown are the means and standard errors (±SE) of three biological replicates. Different letters represent significant differences at p-value < 0.05.

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Fig 7.

Known components of clathrin-mediated endocytosis involved in the RNAi response in WCR.

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Fig 7 Expand