Deletion of RBPJK enhances cell spontaneous differentiation in regular MSC culture.

<p>Mesenchymal stem cells were infected with Control shRNA(Co) or RBPJK shRNA (shRBPJK) lentivirus before being harvested for alkaline phosphatase (ALP) staining, western blot, RT-PCR analysis and BrdU ELISA. (A) Western Blot showed a significant decrease of RBPJK protein expression at day 3 and 14 following shRBPJK lentiviral infection. (B) Fold change in protein level of RBPJK in Western blots was determined by measuring band intensity with ImageJ. (C) Deletion of RBPJK resulted in increased ALP staining at days 3 and 14, compared to controls (Co). Scale bars, 100μm. (D) ALP gene expression was increased at days 3 and 14, with maximal increase at day 14 in RT-PCR analysis. (E) BrdU ELISA to monitor both MSCs proliferation at days 3 and 14 after lentiviral infection. Data are means ± s.d. of three independent experiments performed in duplicate and the control gene expression level at day 3 was set at 1. (*, <i>p</i> < 0.05 compared with control at same time point).