The vitellogenesis-inhibiting hormone (SpVIH) of the crab Scylla paramamosain is not likely to have a vitellogenesis-inhibiting function in the ovary

The final dsRNA was diluted with water to the appropriate concentration (1µg/µL). Primer sequences (dsSpVIH-F/R) are listed in Table 1. The quality of dsVIH was detected by agarose gel electrophoresis (Figure S1).

A female crab’s tissue was placed in one sterile incubation plate, according to Figure S2. Each tissue block was about 50 mg, and each well in the plate contained 2 mL of sterile M199 culture medium and a 1:1000 ratio of prepared ampicillin solution. 10 pmol/mL dsVIH or 3.5 pmol/mL PrVIH was added to wells according to Figure S2, and tissues were collected separately after incubating for 3h at 28 °C. The control group was added with the same volume of 1 × PBS. Each group consisted of five replicates, and the samples were derived from one crab, and 5 crabs were used.