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. 2015 May 1;26(9):1711–1727. doi: 10.1091/mbc.E14-07-1221

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© 2015 Cruse et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License (http://creativecommons.org/licenses/by-nc-sa/3.0).

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FIGURE 1: — MS4A4 in human MC promotes surface KIT receptor expression. (A) RT-PCR for MS4A4 and β-actin in HLMCs. (B) All human MC types tested expressed MS4A4 mRNA. (C) qRT-PCR for MS4A4 mRNA in human MCs. Data calculated as the ratio of MS4A4 compared with β-actin for each sample (n = 3–11). (D) qRT-PCR for MS4A4 mRNA in CD34⁺-derived peripheral blood MC over time during culture (n = 3). (E) qRT-PCR for MS4A4 mRNA in LAD-2 cells using four shRNA constructs targeting MS4A4 (n = 5). (F) Flow cytometry histogram of total MS4A4 protein expression in LAD-2 cells treated with scrambled shRNA control or the shMS4A4v4 construct (isotype, black; scramble, blue; shMS4A4v4, red). (G) Mean flow cytometry data for MS4A4 expression calculated from the geometric MFI and expressed as percentage of scramble control (n = 4). (H) Mean flow cytometry data for surface KIT expression in shMS4A4-transduced LAD-2 cells (n = 4). (I) Flow cytometry histogram of surface KIT expression. (J) Flow cytometry histogram of total KIT expression in permeabilized cells. (K) Flow cytometry histogram of surface CD54 expression. (L) qRT-PCR for KIT mRNA with shMS4A4 constructs. Values are relative expression compared with scramble control (n = 3). (M) Correlation between surface KIT protein expression and relative KIT mRNA expression calculated as relative percentage compared with scramble control. (N) Flow cytometry histograms of annexin V staining from LAD-2 cells transduced with either control shRNA or shMS4A4 constructs at day 7 postinfection. Data are mean + SEM. *p < 0.05, **p < 0.001.