Figure 2.

HuR silencing increases cytoplasmic Rac1 abundance in cultured IEC-6 cells. (A) Representative Western blots in cytoplasmic (left), nuclear (middle), and total (right) Rac1 protein in IEC-6 cells. After cells were transfected with siHuR or C-siRNA, whole-cell lysates were harvested 48 hours thereafter. Equal loading in cytoplasmic, nuclear, and total proteins were monitored by β-tubulin, lamin B, and actin, respectively. (B) Immunostaining of Rac1 in cells described in panel A. Cells were permeabilized and incubated with the anti-Rac1 antibody and then with anti-IgG antibody conjugated with Alexa Fluor. Red arrows point to Rac1 signals. Nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI). Green, signals of Rac1; blue, nuclear stained by DAPI. Scale bar: 5 μm. Three experiments were performed that showed similar results.