Fig. 2.

Suppression of Nrf2 gene expression enhances influenza virus replication. Cultures of differentiated human epithelial cells were infected with lentiviral vectors encoding shRNAs LV-Nrf2 or GFP-scrambled control. At 48 h posttransduction, cells were infected with influenza A/Bangkok/1/79. (A) Total RNA was isolated and subjected to RT-PCR to quantify Influenza HA transcripts and normalized to the expression of β-actin. (B) Apical supernatants collected 24 h postinfection were analyzed for determination of the viral titer indicated in log TCID₅₀. (C) Total cellular lysates were analyzed for Nrf2 and HO-1 protein expression by Western blot. Membrane was stripped and analyzed for β-actin as a loading control. Densitometry was used to quantitate the amounts of protein, and the numbers below the gel indicate the scramble control/experimental sample ratio. Asterisk indicates statistical significance between test sample and control, * P < 0.05.