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. 2010 Apr 28;5(4):e10369. doi: 10.1371/journal.pone.0010369

Figure 9. Separate control of Notch1 transcription by Klf4/Sp3 and p53.

Figure 9

(A) HeLa cells were transfected with siRNAs for Sp3 and Klf4 in parallel with scrambled siRNA controls followed, 48 hours after transfection, by infection with a p53 expressing adenovirus (Adp53) or GFP control (AdGFP), for 24 hours. Levels of Notch1 mRNA expression were determined by real-time RT-PCR. The expected changes of p53, Sp3 and Klf4 expression were also confirmed by real time RT-PCR, with results similar to those shown in previous figures. (B) HeLa cells were transfected with siRNAs for UBE3A, Klf4 and Sp3 alone or in combinations as indicated, in parallel with scrambled siRNA controls. UBE3A and Notch1 expression was assessed by real-time RT-PCR. (C and D) Primary keratinocytes (HKC) (C) and HeLa and SCC13 cells (D were infected with a Klf4 expressing retrovirus or empty vector control for 48 hours, followed by infection with the Adp53 or AdGFP viruses for 24 hours. Notch1 mRNA levels were assessed by real-time RT-PCR.