Da gal4

Fly stocks were raised on standard cornmeal medium and kept at 23 °C, 70% humidity on a 12:12 hours light/dark cycle. Strains used in this study were obtained from Bloomington Drosophila Stock Center (BDSC) and Vienna Drosophila Resource Center (VDRC). Genotypes used in this study were: act5c-gal4 (BDSC 4414), da-gal4 (BDSC 8641), UAS-Ndufs4 RNAi (VDRC 101489), UAS-Bcs1 RNAi (BDSC 51863), UAS-Coa8 RNAi (VDRC 100605). Control strains were obtained in each experiment by crossing the specific gal4 driver line with the genetic background flies w¹¹¹⁸. Coa8 KO flies were generated by Wellgenetics Inc by using CRISPR/Cas9 technology, generating a 676 bp deletion, from the –49th nucleotide relative to ATG to the –34th nucleotide relative to the stop codon of Coa8.

All flies were maintained at 25 °C on standard laboratory medium as described previously^{55 (link)}. The following stocks were obtained from the Bloomington Drosophila stock center (BDSC): w¹¹¹⁸ (wild-type; BDSC 5905), act5c-GAL4 (BDSC 3954), da-GAL4 (BDSC8641), elav-GAL4 (BDSC 458), elav-GAL4; UAS-dcr-2 (BDSC 25750), P{CaryP} attP2 (BDSC 36303), UAS-tra2 RNAi #1 (BDSC 56912), UAS-tra2 RNAi #2 (BDSC 28018), UAS-traF (BDSC 4590), UAS-msl-2 RNAi #1 (BDSC 31627), UAS-msl-2 RNAi #2 (BDSC 35390), UAS-Sxl RNAi #1 (BDSC 34393), UAS-Sxl RNAi #2 (BDSC 38195), Sxl^f7,M1; P{Sxl. + tCa}9A/ + (BDSC 58486), and Sxl^M1,fΔ33/Binsinscy (BDSC 58487). Three gene-switch Gal4 driver lines—act5c-GS-GAL4, S106-GS-GAL4, and 5961-GS-GAL4—were kindly gifted by Dr. Akagi^{56 (link)}.

All flies were maintained at 25 ± 0.5°C and 80 ± 5% humidity under 12 h of light/dark cycles on cornmeal-sucrose-yeast agar media. w¹¹¹⁸ flies were used as wild-type controls. Fly lines were obtained from the Bloomington Drosophila Stock Center (BDSC) and the Vienna Drosophila Resource Center (VDRC). The following stocks were used:
y¹ w^67c23 (BDSC 6599),
da-Gal4 (BDSC 5460),
w¹¹¹⁸ (BDSC 3605),
UAS-wsv056,
tub-GAL80^ts,
CG12116 RNAi (VDRC 6498), and
CG12117 RNAi (VDRC 17017).
Pacific white shrimp L. vannamei of approximately 4–5 g body weight were bought from a commercial farm (Zhanjiang, Guangdong Province, China). Before the experiment, shrimp were acclimated for 1 week in aerated artificial seawater with water salinity, with temperature maintained at 29–31 g/L and 24–26°C. Shrimp were fed with commercial feed three times a day at the rate of 5% body weight. PCR with WSSV-specific primers (Table 1) was performed to ensure that the shrimp were free of WSSV before experiments.