FIG 3.
6-TG treatment inhibits HA and NA glycoprotein processing and accumulation without affecting transcript levels. (A) A549 cells were mock infected or infected with IAV-PR8 at an MOI of 0.1. After 1 h, cell monolayers were washed and overlaid with medium containing drugs at the indicated concentrations or the vehicle control (−). At 24 hpi, cell lysates were collected and analyzed by Western blotting using a polyclonal IAV antibody that detects HA, NP, and M1 as well as antibodies that detect IAV PA, NA, NS1, or cellular actin. A representative blot from 3 independent experiments is shown. (B) The levels of IAV HA and NA mRNAs, and HA and NA genomic vRNAs, were measured by RT-qPCR in A549 cells infected with IAV-PR8 and treated with 2 μM or 10 μM 6-TG or the vehicle control. Total RNA was isolated at 24 hpi. Changes in RNA levels were calculated by the ΔΔCT method and normalized to 18S rRNA. Error bars represent standard deviations (n = 3). Circles represent biological replicates, and lines represent mean values.