FIG. 3.

RNA interference of ALF and WSSV replication in vivo. RNA interference was performed with an injection of 150 μg of ALF dsRNA or GFP dsRNA into crayfish (20 ± 2 g [fresh weight]). The same injection was repeated 24 h after the first dsRNA injection. These crayfish were infected with WSSV by injection of 200 μl of live WSSV stock suspension 12 h after the second dsRNA injection. Hemocyte total RNA was prepared 36 h post-WSSV infection for quantitative RT-PCR. The crayfish ribosomal protein 40s gene was used as an internal control for WSSV VP28 gene and crayfish ALF gene quantitation by quantitative RT-PCR. The experiment has been repeated three times, the data represent means of triplicates, and the error bars indicate standard deviations. Columns 1 and 2 show VP28 and ALF transcript levels, respectively, after injection with GFP dsRNA. Columns 3 and 4 show VP28 and ALF transcript levels, respectively, after injection with ALF dsRNA.