Skip to main content
. Author manuscript; available in PMC: 2016 Mar 5.
Published in final edited form as: Mol Cell Endocrinol. 2015 Jan 14;403:30–38. doi: 10.1016/j.mce.2015.01.015

Fig. 3.

Fig. 3

Twist1 is required for Notch activation-mediated inhibition of MPC chondrogenesis. Limb bud cells in micromass were infected with NICD1 or/and shTwist1 lentivirus for 3 days before being harvested for Western blot, Alcian blue staining and RT-PCR analysis. (A) Twist1 protein levels were significantly reduced by Twist1 shRNA lentiviral infection. (B) Quantification of Twist1 protein expression in Western blots showed 80% inhibition of Twist1 expression when compared to control cells (Co). (C) A significant increase in chondrocyte nodule formation was observed in both shTwist1 infected cells with or without co-infection with NICD1, and a drastic reduction in chondrocyte nodule formation was seen in NICD1-expressing cells. Scale bars, 100 µm. (D) Sox9, type II collagen, and Agc1 gene expression were significantly increased in Twist1 knocking down MPCs with or without overexpressing NICD1. Twist1 expression was significantly increased in NICD1 overexpressing cells and decreased in both Twist1 knocking down MPCs with or without overexpressing NICD1. Data are means ± s.d. of three independent experiments performed in duplicate and the GFP-virus control (Co) gene expression level was set at 1. (*, P < 0.05 compared with control.)