Figure 5
From: Targeting the actin cytoskeleton: selective antitumor action via trapping PKCɛ
PKCɛ overexpression rescues cells from Chondramide A (ChA)-induced apoptosis and noncancerous cells are less susceptible to ChA. (a) MCF7 and MDA-MB-231 cells were transfected with FLAG.PKCɛ or an empty vector and treated with 300 nM ChA for 24 h. The number of dead cells was analyzed using the propidium iodide (PI) exclusion assay for MCF7 cells and Annexin V staining for MDA-MB-231 cells. Overexpression of PKCɛ was confirmed by western blot (insets). The graphs show the results normalized to the according control. Each experiment was independently performed three times. (b) The adenocarcinoma cell lines MCF7, MDA-MB-231 and the nontumorigenic epithelial breast cell line MCF10-A were evaluated for their responsiveness to ChA by monitoring PI-positive cells after 24 h. (c) Comparison of PKCɛ levels is shown in MCF7, MDA-MB-231 and MCF10-A via western blot (graph shows quantification) as well as via immunohistochemistry. GADPH and actin serve as loading control. Each experiment was independently performed three times. (d) Representative PKCɛ tissue stainings of healthy breast tissue and mammary tumor tissue. Nuclei are counterstained with hematoxylin. Bars represent the mean±S.E.M. of three independent experiments performed in triplicates, *P<0.05, **P<0.01, ***P<0.001 (one-way ANOVA, Bonferroni)
