A novel microfluidic flow-cytometry for counting numbers of single-cell β-actins

Lixing Liu , Beiyuan Fan , Hongyu Yang , Deyong Chen , Shuang Zhang , Junbo Wang , Jian Chen
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引用次数: 1

Abstract

As a house keeping protein with stable expressions, β-actin is used as a loading control in normalization of western blotting. However, the actual numbers of β-actins at the single-cell level remain elusive. Based on a home-developed flow cytometry, single-cell numbers of β-actin from 8 cell types (subtypes) and 2 tumour patient samples were quantified as 9.62 ± 4.29 × 105 (A549, Ncell = 14,242), 6.46 ± 3.34 × 105 (Hep G2, Ncell = 35,932), 1.58 ± 0.90 × 106 (MCF 10A, Ncell = 16,650), 1.08 ± 0.48 × 106 (HeLa, Ncell = 26,151), 7.60 ± 4.34 × 105 (PC3, Ncell = 11,922), 1.10 ± 0.72 × 106 (SACC-83, Ncell = 13,616), 8.58 ± 4.54 × 105 (CAL 27, Ncell = 7271), 9.00 ± 4.69 × 105 (CAL 27-LN2, Ncell = 6222), 8.26 ± 4.48 × 105 (Oral Tumour Patient I, Ncell = 359), and 8.19 ± 5.12 × 105 (Oral Tumour Patient II, Ncell = 175), and were analyzed by statistical approaches including one-way analysis of variance, neural network based pattern recognition and Bayesian estimation, with varied expressions of β-actins among different cell types located. The dataset reported in this study may serve as a reference in future studies of quantitative protein analysis.

一种用于计数单细胞β-肌动蛋白的新型微流控流式细胞术
β-肌动蛋白作为一种稳定表达的家养蛋白,在western blotting归一化中被用作负载对照。然而,在单细胞水平上β-肌动蛋白的实际数量仍然难以捉摸。自主开发基于流式细胞术,单细胞的β肌动蛋白从8细胞类型(亚型)和2肿瘤患者样本被量化为9.62±4.29×105 (A549 Ncell = 14242), 6.46±3.34×105(消息灵通的G2, Ncell = 35932), 1.58±0.90×106 (MCF 10, Ncell = 16650), 1.08±0.48×106(海拉,Ncell = 26151), 7.60±4.34×105(生物,Ncell = 11922), 1.10±0.72×106 (sacc - 83, Ncell = 13616), 8.58±4.54×105 (Ncell卡尔27日= 7271),9.00±4.69×105 (CAL 27-LN2 Ncell = 6222),分别为8.26±4.48 × 105(口腔肿瘤患者I, Ncell = 359)和8.19±5.12 × 105(口腔肿瘤患者II, Ncell = 175),采用单因素方差分析、基于神经网络的模式识别和贝叶斯估计等统计方法进行分析,发现β-肌动蛋白在不同细胞类型中的表达存在差异。本研究的数据集可为今后蛋白质定量分析的研究提供参考。
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