|
Fig. 3
Decrease of Cav-1b expression negatively affects the IFN pathway.
A) Stat1 gene expression was assessed by qRT-PCR in Control MO and Cav-1b MO embryos that were either SHRV infected or uninfected. Total RNA was extracted from 10 fish per treatment, cDNA synthesized and Stat1 mRNA expression assessed by qRT-PCR 24 hpi. The data are representative of three individual experiments and error bars indicate SEM. Each bar represents the mean fold induction of SHRV-infected embryos over corresponding controls. All expression values were normalized to zebrafish 18s. B) ISRE promoter activity is dampened in Cav-1b knockdown ZFL cells upon SHRV infection. ZFL cells were transfected with 250 ng of zISRE-luc construct along with 250 ng of cav-1b MO or control MO. Twenty four hours post transfection the ZFL cells were infected with SHRV at an MOI of 0.01. Cells were harvested for luciferase measurements 24 hpi. The graph shows relative luminescence units of control uninfected cells compared to cav-1b MO or control infected cells. Error bars are representative of SEM for two experiments. (**, p<0.001).