Fig. 3

Efficiency of Notch-1 knockdown in ACC-M cells by lentiviral vectors bearing shRNA. ACC-M cells were infected with lentiviral vectors pLenR-Notch-1-shRNA1, 2, and 3 targeting Notch-1 or scrambled shRNA vector (ACC-Mock (PC)) as detailed in Materials and methods. The Notch-1 mRNA transcript levels were determined by quantitative RT-PCR (a) and Notch-1 protein expression by Western blotting assays (b). Data shown in (c) are expressed as mean ± SD of at least three independent experiments in triplicate. Representative blots are shown in (d) of at least three independent experiments. Notch-1 protein expression was normalized against GAPDH. *p < 0.01 compared with non-infected or scrambled control ACC-M cells