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. Author manuscript; available in PMC: 2012 Aug 1.
Published in final edited form as: Mol Cancer Res. 2011 Jun 21;9(8):1126–1138. doi: 10.1158/1541-7786.MCR-11-0162

Figure 5. Association of reduced methylation in the FILIP1L promoter with restoration of FILIP1L expression in ovarian cancer cells following treatment with a DNA demethylating agent.

Figure 5

A, Real-time RT-PCR analysis for FILIP1L on cDNA from ES2, OVCA429, SKOV3 and OVCAR8 cells treated with either 5-aza-2′-deoxycytidine (DAC) or Trichostatin A (TSA). The y axis represents a fold change over DMSO-treated control cells where each value was standardized with the housekeeping gene hRPL7. Error bars indicate SEM (n = 3). P value comparison between DMSO-treated control and either DAC- or TSA-treated experiments were: both 0.1 and 1 μM DAC (p<0.0001) and both 0.1 and 1 μM TSA (not significant (NS)) for ES2; both 10 and 25 μM DAC (p<0.0001) and both 0.3 and 1 μM TSA (NS) for OVCA429; 1 μM DAC (p=0.0002), 3 μM DAC (p=0.0004) and both 0.3 and 1 μM TSA (NS) for SKOV3; 0.1 μM DAC, 1 μM DAC and 0.1 μM TSA (NS), and 1 μM TSA (p=0.0003) for OVCAR8. The result is an average of three independent experiments. B, Immunoblot analysis for FILIP1L in the same cells used in section A. GAPDH blot is shown as the loading control. Note that in FILIP1L-high expressing OVCAR8 cells, DAC treatment did not result in increased FILIP1L expression whereas TSA treatment decreased FILIP1L expression. The result is representative of three independent experiments. C, DNA methylation status of the CpG island in the FILIP1L promoter from the same cells used in section A was analyzed by Sequenom® EpiTYPER Mass Array. Mass Array results are shown as described in Figure 4B. The result is an average of three independent experiments. D, Mass Array results are shown as described in Figure 4C from the same cells used in section A. P value comparison between DMSO-treated control and DAC- or TSA-treated experiments were: 1 μM DAC (p<0.0001) and 1 μM TSA (NS) for ES2; 25 μM DAC (p=0.0009) for OVCA429; 3 μM DAC (p<0.0001) for SKOV3; 1 μM DAC (NS) for OVCAR8. Note that either TSA-treated ES2 or DAC-treated OVCAR8 did not decrease DNA methylation compared with DMSO-treated control.