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. 2017 Aug 21;7:8952. doi: 10.1038/s41598-017-08357-4

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Arp2/3 complex promoted branched actin polymerization is required for surface TCR maintainance via regulating TCR⁺ endosome trafficking. (a) Flow cytometry analysis of surface and total TCRβ levels. (b) Histogram showing the MFI of surface (n = 13) and total TCRβ (n = 10) levels. (c) Control or Arpc2 KD Jurkat T cells were stained with anti-TCRβ-647 on ice after they were crosslinked at 37 °C for the indicated times. The cells were then stripped and analyzed for TCR internalization using FACS (n = 5). (d) Flow cytometry analysis of internalized TCR recycling in control or Arpc2 KD Jurkat T cells. The data are presented the percentage of internalized TCR receptors that had recycled back to the cell surface. (n = 3). (e) Immunofluorescence analysis of the location of CD3e (greeen), Arpc2 (red) and EEA1/Rab5 (blue) in Arpc2-mCherry Jurkat T cells. Bar is 5 μm. At least 30 cells were analyzed and representative images are shown. (f) Flow cytometry and (g) MFI analysis of TCR surface (n = 8) and total (n = 8) level in cytD treated Jurkat T cells compared with control cells. (h) Control or cytD treated Jurkat T cells were stained with anti-TCRβ-647 on ice, after crosslinking at 37 °C for the indicated times, cells were stripping and analyzed for TCR internalization by FACS. (n = 5). (i) Flow cytometry analysis of internalized TCR recycling in control or cytD treated Jurkat T cells. The data are presented as the percentage of internalized TCR receptors that have recycled back to the cell surface. (n = 3). The data are means ± S.D., for all panels: *P < 0.05; ***P < 0.001 by Student’s t-test, N.S.: no significance. All results are representative from at least three independent experiments.