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. 2006 Aug 31;34(16):4527–4536. doi: 10.1093/nar/gkl628

Figure 3.

Figure 3

Knockdown of Plk1 mRNA and protein by Tet-inducible expression of shRNA/Plk1 using different promoter types stably integrated into the genome of HeLa cell clones. (A) Northern blot analysis of Plk1 mRNA. HeLa cell clones containing a wild-type H1 promoter constitutively expressing shRNA coding for a mismatch sequence (MM) was used as control (lane 1); Plk1 mRNA expression in Tet-inducible US-type promoter-derived cell clones driving the expression of shRNA/Plk1 (lanes 2–5); Plk1 mRNA expression in Tet-inducible DS-type promoter-derived cell clones driving the expression of shRNA/Plk1 (lanes 6–9); Plk1 mRNA expression in Tet-inducible US/DS-type promoter-derived cell clones driving the expression of shRNA/Plk1 (lanes 10–13). Plk1 mRNA expression in Tet-inducible US-, DS- and US/DS-type promoter-derived cell clones driving the expression of shRNA/Plk1MM (14–22). Administration of different Dox concentrations lasted for 120 h. For standardization ethidiumbromide staining of 28S rRNA was chosen. Plk1 mRNA levels were presented as percentage of Plk1 mRNA levels in cell clones constitutively expressing mismatched shRNA. (B) Immunoblot analysis of Plk1 protein expression. Same distribution of samples as shown in (A). For standardization β-actin expression was used. Plk1 protein levels were presented as percentage of Plk1 protein levels in cell clones constitutively expressing mismatched shRNA. Significant reductions (P < 0.05) are indicated with an asterisk.