FIG 6.

The level of iron ions bound by ferritin decreased after WSSV infection. (A) Stomachs excised from L. vannamei shrimps 0, 2, 4, 8, 12, and 24 h after injection with PBS (upper panel) or WSSV (lower panel) were homogenized, and the tissue lysates were mixed with anti-Lvferritin antibody coupled with protein A agarose. The immunoprecipitation products were analyzed by Western blotting to evaluate the amount of total ferritin and by AAS to determine the quantity of iron ions bound by ferritin. (B) The ratio of iron concentration to total ferritin was calculated, and the initial value was set to 1.0 for both the PBS and WSSV injection groups. (C) Stomach tissue lysates prepared from PBS-injected (top) or WSSV-injected (bottom) shrimps were also subjected to Western blot analysis using the anti-Lvferritin antibody or anti-actin antibody to evaluate ferritin expression levels. (D) The ratio of total ferritin to actin was calculated, and the initial value was set to 1.0 for both the PBS and WSSV injection groups. Each bar represents the mean ± SD for 3 samples, where each sample was pooled from 3 shrimps. Unpaired Student's t tests were used to test for statistically significant differences (*, P < 0.05).