This work is dedicated to the memory of Elizabeth D. Hay and her long-standing interest in EMT in development and tumor genesis.
Special Issue Research Article
The role of twist during palate development†
Article first published online: 12 AUG 2008
Copyright © 2008 Wiley-Liss, Inc.
Special Issue: Special Focus on the Extracellular Matrix, in Memory of Dr. Elizabeth D. Hay
Volume 237, Issue 10, pages 2716–2725, October 2008
How to Cite
Yu, W., Kamara, H. and Svoboda, K. K. H. (2008), The role of twist during palate development. Dev. Dyn., 237: 2716–2725. doi: 10.1002/dvdy.21627
- Issue published online: 24 SEP 2008
- Article first published online: 12 AUG 2008
- Manuscript Accepted: 20 MAY 2008
- Baylor Oral Health Foundation
- March of Dimes. Grant Number: 6-FY06-321
The Supplementary Material referred to in this article can be viewed online.
|SuppFigs1-3.tif||18542K||Suppl. Fig. 1. Immunofluorescent staining of Twist protein in vitro. After culturing for 24 hr, the Twist protein (green) was expressed in the epithelial layer where the MES would have formed. Nuclei were counterstained red. Scale bar = 37.5 μm. Suppl. Fig. 2. siRNA transfection efficiency in organ culture. The siRNA transfection efficiency was monitored by using a fluorescent oligo (Block-iT, Invitrogen). The fluorescein-labeled oligo was transfected to the tissue at a concentration of 200 nM and viewed with the confocal microscope as whole mount after 24 hr. A filter (488 nm) was used for detecting the oligo. Multiple optical sections were merged into a projected image. Scale bar = 75 μm. Suppl. Fig. 3. Western blot of Twist protein expression after siRNA culture. Whole palatal shelves were lysed in the RIPA buffer (RadioImmunoPrecipitation Assay Buffer, Sigma) and used for Western blots. The concentrations of total protein in each sample were determined by BCA assay (Pierce). Ten micrograms total protein was loaded in each well on a 4–12% NuPage Bis-Tris gel (Invitrogen). Protein was transferred to PVDF membrane (Millipore). The membrane was incubated with polyclonal primary antibody against Twist (1:1,000, Santa Cruz). Secondary antibodies conjugated with alkaline phosphatase were used at 1:1,000, visualized with the chemiluminescence (Invitrogen), and captured with the Kodak Image station 440<INF>CF</INF> (Kodak Digital Science). A single band at MW about 28 kDa was detected. Positive control (HeLa nuclear extract) migrated at the same mw as experimental samples.|
Please note: Wiley Blackwell is not responsible for the content or functionality of any supporting information supplied by the authors. Any queries (other than missing content) should be directed to the corresponding author for the article.