Fig 4.

(A and B) Effects of NH₄Cl (A) and chloroquine (CQ) (B) on ISKNV infection. Cells were pretreated for 1 h with various concentrations of reagents as indicated or were left untreated (as a positive control), and ISKNV was then added and incubated for 4 h. After 72 h of incubation, the cells were processed for IFA or WB with anti-ORF101L antibody. For the IFA, the viral infections were quantified as the percentage of positive, treated cells relative to the number of untreated control cells. The viral infection rate of cells not treated with reagents (as a positive control) was arbitrarily set as 100%. The data shown are the means and standard deviations of the results from three independent experiments. For WB, endogenous β-tubulin was included as an internal loading control. Lanes marked “+” indicate untreated control cells, and lanes marked “−” indicate the negative controls without ISKNV infection. (C) Internalization of AF488-LDL into MFF-1 cells treated with lysosomotropic agents. Cells were mock treated (no drugs) (a) or pretreated with NH₄Cl (50 mM) (b) or CQ (50 μM) (c), followed by incubation with AF488-LDL for 30 min.