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Absolute Copy Numbers of β-Actin Proteins Collected from 10,000 Single Cells
Semi-quantitative studies have located varied expressions of β-actin proteins at the population level, questioning their roles as internal controls in western blots, while the absolute copy numbers of β-actins at the single-cell level are missing. In this study, a polymeric microfluidic flow cytomet...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6187317/ https://www.ncbi.nlm.nih.gov/pubmed/30424187 http://dx.doi.org/10.3390/mi9050254 |
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author | Fan, Beiyuan Li, Xiufeng Liu, Lixing Chen, Deyong Cao, Shanshan Men, Dong Wang, Junbo Chen, Jian |
author_facet | Fan, Beiyuan Li, Xiufeng Liu, Lixing Chen, Deyong Cao, Shanshan Men, Dong Wang, Junbo Chen, Jian |
author_sort | Fan, Beiyuan |
collection | PubMed |
description | Semi-quantitative studies have located varied expressions of β-actin proteins at the population level, questioning their roles as internal controls in western blots, while the absolute copy numbers of β-actins at the single-cell level are missing. In this study, a polymeric microfluidic flow cytometry was used for single-cell analysis, and the absolute copy numbers of single-cell β-actin proteins were quantified as 9.9 ± 4.6 × 10(5), 6.8 ± 4.0 × 10(5) and 11.0 ± 5.5 × 10(5) per cell for A549 (n(cell) = 14,754), Hep G2 (n(cell) = 36,949), and HeLa (n(cell) = 24,383), respectively. High coefficients of variation (~50%) and high quartile coefficients of dispersion (~30%) were located, indicating significant variations of β-actin proteins within the same cell type. Low p values (≪0.01) and high classification rates based on neural network (~70%) were quantified among A549, Hep G2 and HeLa cells, suggesting expression differences of β-actin proteins among three cell types. In summary, the results reported here indicate significant variations of β-actin proteins within the same cell type from cell to cell, and significant expression differences of β-actin proteins among different cell types, strongly questioning the properties of using β-actin proteins as internal controls in western blots. |
format | Online Article Text |
id | pubmed-6187317 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-61873172018-11-01 Absolute Copy Numbers of β-Actin Proteins Collected from 10,000 Single Cells Fan, Beiyuan Li, Xiufeng Liu, Lixing Chen, Deyong Cao, Shanshan Men, Dong Wang, Junbo Chen, Jian Micromachines (Basel) Article Semi-quantitative studies have located varied expressions of β-actin proteins at the population level, questioning their roles as internal controls in western blots, while the absolute copy numbers of β-actins at the single-cell level are missing. In this study, a polymeric microfluidic flow cytometry was used for single-cell analysis, and the absolute copy numbers of single-cell β-actin proteins were quantified as 9.9 ± 4.6 × 10(5), 6.8 ± 4.0 × 10(5) and 11.0 ± 5.5 × 10(5) per cell for A549 (n(cell) = 14,754), Hep G2 (n(cell) = 36,949), and HeLa (n(cell) = 24,383), respectively. High coefficients of variation (~50%) and high quartile coefficients of dispersion (~30%) were located, indicating significant variations of β-actin proteins within the same cell type. Low p values (≪0.01) and high classification rates based on neural network (~70%) were quantified among A549, Hep G2 and HeLa cells, suggesting expression differences of β-actin proteins among three cell types. In summary, the results reported here indicate significant variations of β-actin proteins within the same cell type from cell to cell, and significant expression differences of β-actin proteins among different cell types, strongly questioning the properties of using β-actin proteins as internal controls in western blots. MDPI 2018-05-22 /pmc/articles/PMC6187317/ /pubmed/30424187 http://dx.doi.org/10.3390/mi9050254 Text en © 2018 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Fan, Beiyuan Li, Xiufeng Liu, Lixing Chen, Deyong Cao, Shanshan Men, Dong Wang, Junbo Chen, Jian Absolute Copy Numbers of β-Actin Proteins Collected from 10,000 Single Cells |
title | Absolute Copy Numbers of β-Actin Proteins Collected from 10,000 Single Cells |
title_full | Absolute Copy Numbers of β-Actin Proteins Collected from 10,000 Single Cells |
title_fullStr | Absolute Copy Numbers of β-Actin Proteins Collected from 10,000 Single Cells |
title_full_unstemmed | Absolute Copy Numbers of β-Actin Proteins Collected from 10,000 Single Cells |
title_short | Absolute Copy Numbers of β-Actin Proteins Collected from 10,000 Single Cells |
title_sort | absolute copy numbers of β-actin proteins collected from 10,000 single cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6187317/ https://www.ncbi.nlm.nih.gov/pubmed/30424187 http://dx.doi.org/10.3390/mi9050254 |
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