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. 2003 Aug 1;31(15):e77. doi: 10.1093/nar/gng077

Figure 2.

Figure 2

Western blot analysis of DNMT1 expression in cells transfected with various vectors. Vectors transfected into HCT116 cells are indicated with the symbols used in Figure 1. As a control, expression of β-actin and PCNA is shown. (A) Suppression of DNMT1 by siRNA targeting DNMT1 transcribed in vivo under non-selective conditions. (B) Suppression of DNMT1 by hairpin siRNA with inducible vectors. DNMT1 expression in HCT116 cells transfected with the plasmids indicated with or without treatment with doxycycline is shown. (C) DNMT1 knockdown with the inducible vector and selection marker. Expression of DNMT1, DNMT3A and DNMT3B was determined by western blot analysis. HCT116 cells transfected with the various plasmids indicated with or without doxycycline treatment are shown. Western blot analysis showed that DNMT1 levels were decreased by addition of doxycycline in a dose-dependent fashion, with complete suppression at 10 µg/ml. Expression of DNMT3A and DNMT3B was not affected by 10 µg/ml of doxycycline, showing the specificity of the knockdown.