Figure 5
Genistein interferes with HIV-mediated actin dynamics in resting memory CD4 T cells. (A) Resting memory CD4 T cells from two donors were pretreated with genistein (3.7 μM) or mock-treated for 1 hour at 37°C, and then stimulated with HIV-1NL4-3 (100 ng) for various times. Cells were fixed, stained with FITC-phalloidin for F-actin, and then analyzed with a flow cytometer. (B) Confocal images of cells stained in (A), showing F-actin staining. Images were acquired and analyzed at identical conditions for all samples. (C) Resting memory CD4 T cells were pretreated with 10 μM genistein for 1 hour, and then treated with HIV-1NL4-3 for a time course. Cells were then analyzed by Western blot for LIMK and cofilin activation, using an anti-phospho-LIMK antibody or an anti-cofilin antibody. The same blots were also probed with an anti-human GAPDH antibody for loading control and normalization.