Western Blot Analysis of pBEC Proteins

pBECs were lysed in RIPA buffer, and all proteins were standardized to 5 μg and were resolved by SDS-PAGE, and transferred onto nitrocellulose membranes for detection of CD147, histone deacetylase (HDAC) 4, MMP-9, c-Myc and SP1 (Abcam, UK), in cell lysates. Glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was detected as a loading control for proteins in cell lysates [12 (link), 13 (link)]. Protein estimation was determined by densitometry, using Image lab software (version 4.1), and values were expressed as protein/GAPDH ratio and normalized to media controls at baseline from pBEC of non-asthmatics. All antibodies used in this study are commercially available and have been validated by the source company.

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Moheimani F., Koops J., Williams T., Reid A.T., Hansbro P.M., Wark P.A, & Knight D.A. (2018). Influenza A virus infection dysregulates the expression of microRNA-22 and its targets; CD147 and HDAC4, in epithelium of asthmatics. Respiratory Research, 19, 145.

Publication 2018

CD147 MMP-9 c-Myc

Antibodies Asthmatics Buffer C myc Cd147 Cell Densitometry Glyceraldehyde 3 phosphate dehydrogenase Histone deacetylase 4 Membranes Mmp 9 Nitrocellulose Protein Ripa Sds page

Corresponding Organization :

Other organizations : University of Newcastle Australia

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Variable analysis

independent variables

Protein samples from pBECs

dependent variables

Levels of CD147
Levels of HDAC4
Levels of MMP-9
Levels of c-Myc
Levels of SP1

control variables

Protein samples were standardized to 5 μg
GAPDH was detected as a loading control for proteins in cell lysates

positive controls

Protein estimation was determined by densitometry, using Image lab software (version 4.1)

negative controls

Protein/GAPDH ratio was normalized to media controls at baseline from pBEC of non-asthmatics

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