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https://athenacommons.muw.edu/urc/2021/stem/3
Department
Sciences & Mathematics
Format of Presentation
Oral Presentation
Research Category
STEM
Description
Endogenous CFTR Expression in Human Pancreatic Cell Lines
Zithlay Amezquita, Justin Labonte, Ghanshyam Heda
CFTR (cystic fibrosis transmembrane conductance regulator) is a plasma membrane protein that function as a chloride ion channel in epithelial cells. Mutation in this protein affects its function targeting many organs and result in cystic fibrosis (CF). CF is a genetic disease that is most common among Caucasians of northern European origin. Detection of CFTR at the endogenous levels is cumbersome. The goal of this project therefore is to detect the expression of CFTR in cell lines that are not transfected. Cell lines expressing mutated CFTR (CFPAC) and wild-type (Capan-1) were treated with 5mM SB for 60 hrs to upregulate the CFTR expression. Exogenous CFTR expressing cell line (CFBE-wt) and its parental cell line (CFBE410) were include as a positive and negative controls. Endogenous CFTR protein expression in these cell lines can be visualized by western blotting and quantitated. CFTR mRNA expression was also detected by a qPCR analysis and compared with their expression levels with the exogenous CFTR expression. This is a first report of detection of endogenous CFTR protein expression and was possible by using a newly formulated blocking buffer [Heda et al, BioTechniques, 68(6), 319-325, 2020].
Recommended Citation
Amezquita, Zithlaly and Labonte, Justin Taylor, "Endogenous CFTR Expression in Human Pancreatic Cell Lines" (2021). Undergraduate Research Conference. 3.
https://athenacommons.muw.edu/urc/2021/stem/3
Endogenous CFTR Expression in Human Pancreatic Cell Lines
Endogenous CFTR Expression in Human Pancreatic Cell Lines
Zithlay Amezquita, Justin Labonte, Ghanshyam Heda
CFTR (cystic fibrosis transmembrane conductance regulator) is a plasma membrane protein that function as a chloride ion channel in epithelial cells. Mutation in this protein affects its function targeting many organs and result in cystic fibrosis (CF). CF is a genetic disease that is most common among Caucasians of northern European origin. Detection of CFTR at the endogenous levels is cumbersome. The goal of this project therefore is to detect the expression of CFTR in cell lines that are not transfected. Cell lines expressing mutated CFTR (CFPAC) and wild-type (Capan-1) were treated with 5mM SB for 60 hrs to upregulate the CFTR expression. Exogenous CFTR expressing cell line (CFBE-wt) and its parental cell line (CFBE410) were include as a positive and negative controls. Endogenous CFTR protein expression in these cell lines can be visualized by western blotting and quantitated. CFTR mRNA expression was also detected by a qPCR analysis and compared with their expression levels with the exogenous CFTR expression. This is a first report of detection of endogenous CFTR protein expression and was possible by using a newly formulated blocking buffer [Heda et al, BioTechniques, 68(6), 319-325, 2020].
https://athenacommons.muw.edu/urc/2021/stem/3