FIG 2.

Effects of RSV infection on HMGB1 protein levels in A549 cells. (A) A549 cells were mock infected or infected with RSV at an MOI of 1.0, and the viability of the cells was evaluated by Trypan blue exclusion. (B) Total cell lysates and cytoplasmic and nuclear extracts prepared from uninfected or RSV-infected A549 cells for 6, 15, 24, and 48 h p.i. were resolved on 12% SDS-PAGE, and Western blotting was performed using antibody against HMGB1 protein. Membranes were stripped and reprobed for β-actin as an internal control for protein integrity and loading for total lysates and cytoplasmic extracts, and lamin B was used for nuclear extracts. (C to E) Densitometric analyses of HMGB1 Western blot band intensity for total cell lysates (C), cytoplasmic extracts (D), and nuclear extracts (E). Bands in RSV-infected samples were normalized to the uninfected sample background for each time point. Open bars, control; solid bars, RSV. The figure is representative of three independent experiments. P values were <0.05 (*), <0.005 (**), and <0.0005 (***) compared with uninfected cells.