FIG 9.

Effect of siRNA-mediated knockdown of EqCXCL16 on EAV infection. (A) Stable HEK-EqCXCL16 cells were transfected with two different siRNAs (siRNA1[row c] and siRNA2 [row d]) directed against EqCXCL16 mRNA. Mock-transfected cells (row a) or cells transfected with scrambled siRNA (row b) were considered negative controls. At 30 h posttransfection, cells were infected with EAV sVBSmCherry at an MOI of 1.0 for 18 h. Cells were fixed, stained with Gp anti-EqCXCL16 pAb, and analyzed using an inverted fluorescence microscope for the evaluation of mCherry and EqCXCL16 expression. Green staining reflects the surface expression of EqCXCL16, while red staining indicates mCherry expression. In the presence of EqCXCL16 (rows a and b), mCherry expression can be visualized. In contrast, when cells were transfected with siRNAs, mCherry expression was suppressed (rows c and d). (B) Stable HEK-EqCXCL16 cells were transfected with siRNA1, and at 30 h posttransfection, the cells were infected with EAV sVBSmCherry. After 18 h of incubation, the cells were lysed and analyzed by a WB assay using nsp-1 (a), β-actin (b), and rabbit anti-EqCXCL16 (c) Abs. Arrows, locations of nsp-1 (a), β-actin (b), and EqCXCL16 (c).