Figure 5

NDV infection reduces β-catenin expression. TOPFlash/FOPFlash luciferase assay was performed in SAS cells infected with NDV (0.1MOI). LiCl at a concentration of 20 mM was used to stimulate the cells along with mock infected and NDV infected cells controls. TOP/FOP ratio is plotted on Y axis and was broken at 3 and restarted at 30 to accommodate the higher values. The dashed line states the basal value corresponding to mock treated samples. The data is average of three independent experiments (A). The mRNA levels of various β-catenin regulated genes were analyzed 48 hr post-NDV infection, the experiment was performed at least three times and GAPDH was used as a normalizing control (B). Data are represented as fold change upon NDV infection relative to control. Western blots showing the suppression of β-catenin beyond detection and down regulation of β-catenin regulated genes 48 hr post-infection (C). SAS cell lysates were collected at 24, 48 and 72 hr post-NDV (0.1MOI) infection and analyzed for p-Akt (Ser473), total GSK-3β, p-GSK-3β (Ser9), β-catenin, and MMP-7 expression by western blot (D). The β-actin serves as the loading control. Cytoplasmic and nuclear fractions were collected from SAS cells 48 hr post-NDV infection (0.1MOI) and analyzed for β-catenin by western blot. The β-actin and histone serve as the loading controls (E). T-test using Microsoft Excel, *P < 0.05, **P ≤ 0.01, ***P ≤ 0.001.