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Inhibition of Notch pathway by Notch siRNA or GSI inhibited Rink-1 cell growth.

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posted on 20.02.2013, 17:50 authored by Zhiwei Wang, Sanjeev Banerjee, Aamir Ahmad, Yiwei Li, Asfar S. Azmi, Jason R. Gunn, Dejuan Kong, Bin Bao, Shadan Ali, Jiankun Gao, Ramzi M. Mohammad, Lucio Miele, Murray Korc, Fazlul H. Sarkar

A, Left panel, Inhibition of Rink-1 cell growth by Notch 1–4 siRNA tested by MTT assay. The results were plotted as means ± SD of three separate experiments having six determinations per experiment for each experimental condition. Middle and Right panel: L-685,458 and DAPT were γ-secretase inhibitors (GSI), which prevent the cleavage of the Notch receptor, blocking Notch signal transduction. GSI significantly inhibited Rink-1 cell growth. Cells were seeded in 96-well plates at 5,000 cells per well and treated with GSI for 72 hours. After treatment, cell densities were determined by MTT assay. Each value represents the mean ± SD (n = 6) of three independent experiments. *P<0.05, compared to the control. B, The expression of Notch pathway was down-regulated in Rink-1 cells treated with GSI or transfected with Notch 1–4 siRNA as assessed by western blotting analysis. C, The expression of Notch target genes was down-regulated in Rink-1 cells treated with GSI or transfected with Notch-2 siRNA or Notch-4 siRNA as assessed by as assessed by real-time RT-PCR.

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