Figure 5

Time course response of beclin-1 expression (A), and inhibition of beclin-1 induction by astragalin in H₂O₂-stimulated BEAS-2B cells (B) and in OVA-challenged mouse airways (C). BEAS-2B cells were cultured with 20 μM H₂O₂ in the absence and presence of 1–20 μM astragalin for up to 24 h. Cell lysates were prepared for Western blotting with a primary antibody against beclin-1 (A and B). β-Actin protein was used as an internal control. The bar graphs (mean ± SEM, n = 3) represent quantitative results of the upper bands obtained from a densitometer. Means without a common letter differ, P < 0.05. Immunofluorescence analysis showing inhibition of beclin-1 induction by astragalin in OVA-challenged mouse lung tissues (C). Beclin-1 indentified as red staining was visualized with a Cy3-conjugated secondary antibody and nuclear staining was done with 4′,6-diamidino-2-phenylindole. Each photograph is representative of four mice. Magnification: 200-fold.