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. Author manuscript; available in PMC: 2012 Aug 1.
Published in final edited form as: Cancer Res. 2011 Dec 22;72(3):810–820. doi: 10.1158/0008-5472.CAN-11-1052

Figure 5. SMYD3 knockdown causes epigenetic repression of MMP-9 in human fibrosarcoma cells.

Figure 5

A and B) HT1080 cells were transfected with two different siRNA oligonucleotides against SMYD3 or control followed by RTqPCR analysis of SMYD3 (A) and MMP-9 expression (B), 48 hours after transfection. GAPDH mRNA was used for normalization.

C) Western analysis in HT1080 cells transfected with two different siRNA oligonucleotides against SMYD3 or control siRNA, showing SMYD3 and MMP-9 proteins. β-actin was used as control. MW=molecular weight marker.

D) Luciferase activity in HT1080-2.2 cells with an integrated MMP-9 promoter-Luciferase reporter, following transfection with siRNA oligonucleotides against SMYD3 or control siRNA (Ctrl).

E) Amplicon surrounding putative SMYD3 binding sites (ChIP-1H) in the human MMP-9 promoter. The right arrow marks the transcription start site.

F) Chromatin Immunoprecipitation was performed with antibodies recognizing SMYD3, H3K4me3 or Acetylated-H3. GAPDH promoter is a positive control for the histone marks, and the Afamin promoter a negative control.

*p<0.05, **p<0.01, ***p<0.001