Rottlerin stimulates apoptosis in NHEKs.

<p>(A) NHEKs were treated with various concentrations of rottlerin for 48 h. Apoptotic cell-death was measured by annexin V-PE/7AAD staining in NHEKs. The graph represents the mean±SEM of the percent of apoptotic cells in the flow cytometry results. The data are presented as the mean±SEM (n = 3). *<i>P</i><0.05, **<i>P</i><0.01 vs. control. The data are representative of three independent experiments. (B) Immunofluorescence analysis of p53 in NHEKs after treatment with 5μM rottlerin for 24h. The cells were stained with an anti-p53 antibody, and revealed with cy3-conjugated secondary antibody and DAPI (blue). The arrows point out that p53 translocate from the cytoplasm to the nucleus. Scale Bar = 20 μm. The images are representative of three independent experiments. (C) Western blotting analysis of Bax, Bcl2, cleaved Caspase-3 protein expression after NHEKs treated with 1, 5, and 10μM rottlerin for 48h. GAPDH was used as the loading control. The data are representative of three independent experiments.