Identification of a Calcitriol-Regulated Sp-1 Site in the Promoter of Human CD14 using a Combined Western Blotting Electrophoresis Mobility Shift Assay (WEMSA)

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Author: Moeenrezakhanlou, A
Author: Reiner, NE
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Methods Article

Identification of a Calcitriol-Regulated Sp-1 Site in the Promoter of Human CD14 using a Combined Western Blotting Electrophoresis Mobility Shift Assay (WEMSA)

Alireza Moeenrezakhanlou¹, Devki Nandan² and Neil E. Reiner²^*

¹ Department of Microbiology and Immunology, University of British Columbia, Faculties of Medicine and Science, and Vancouver Coastal Health Research Institute (VCHRI), Vancouver, British Columbia, Canada, V5Z 3J5 and School of Medicine, Hamadan University of Medical Sciences, Hamadan, Iran.
² Department of Microbiology and Immunology, University of British Columbia, Faculties of Medicine and Science, and Vancouver coastal health Research Institute (VCHRI), Vancouver, British Columbian, Canada, V5Z 3J5.

* To whom correspondence should be addressed: Neil E. Reiner, Department of Microbiology and Immunology, University of British Columbia, Faculties of Medicine and Science, and Vancouver Coastal Health Research Institute (VCHRI). Division of Infectious Diseases, University of British Columbia, Rm 452D, 2733 Heather Street, Vancouver, BC. Canada. Phone: 604-875-4011. Fax: 604-875-4013. Email: ethan@interchange.ubc.ca

Biol. Proced. Online 2008;10:29-35. doi:10.1251/bpo140
Submitted: March 23, 2007; Accepted: December 12, 2007; Published: February 17, 2008.

Indexing terms: Electrophoretic Mobility Shift Assay; Transcription Factors; Antigens, CD14.

Abbreviations: TF, transcription factor; VDRE, vitamin D response element; WEMSA, western blot electrophoretic mobility shift assay.

Figure 1 Enlarged

Fig. 1:

CD14 proximal promwter. This region spanning 469 bp upstream of the transcriptional start site has been shown to be critical for the induction of CD14 transcription in response to calcitriol. Putative Sp-1-like transcription factor-binding sites in this region were identified using the TFSEARCH program (http://www.cbrc.jp/research/db/TFSEARCH.html) and are highlighted in boldface, underlined and labeled as (1) 5'-GGGGGGTTGG-3' at position -345 to -336, (2) 5'-GTCCCTCCCCCT-3' at position -159 to -148, (3) 5'-AGGGGGCTGGC-3' at position -113 to -100 and (4) 5'-AGAGGTGGGGAGG-3' at position -91 to -79.

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