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. 2012 Jun 29;7(6):e40355. doi: 10.1371/journal.pone.0040355

Figure 5. Effect of cryptopleurine on the TNF-α-induced NF-κB-dependent inflammatory cytokines, antiapoptotic, and proliferation genes expression.

Figure 5

(A) MDA-MB231 cells were preincubated with indicated concentrations of cryptopleurine for 12 h and then treated with TNF-α (20 ng/ml) for an additional 12 h. RNA was isolated from cells, reverse-transcribed, and analyzed by real-time PCR for IL-6, IL-8, and IL-1β. Data represented as mean ± standard deviation of three independent experiments. *p<0.05, ***p<0.001, significantly different when compared with TNF-α-stimulated normal cells. (B) MDA-MB231 cells were incubated with 30 nM cryptopleurine for 12 h and then incubated with 20 ng/ml TNF-α for the indicated times. Whole cell extracts were analyzed by Western blot using indicated antibodies for TRAF2, Bcl2, cIAP1, FLIP, COX-2, cyclinD1, and tubulin. (C) MDA-MB231, MDA-MB435, RAW264.7, and Hep3B cells were plated in triplicate, treated with 0, 10 or 30 nM cryptopleurine, and subjected to MTT assay on days 1, 2, 3 to analyze cell proliferation. Absorbance was measured at 570 nm.