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. 2017 Jun 15;28(12):1688–1700. doi: 10.1091/mbc.E17-02-0128

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© 2017 Chehab, Santos, 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 3: — Munc13-4 localizes to WPBs via its MHD1/MHD2 region. TIRF images of live HUVECs expressing different truncated versions of YFP–Munc13-4 and VWF-RFP as WPB marker for 48 h. Right, domain structures of the respective Munc13-4 mutants lacking the C2A (aa 109–284), C2B (aa 904–1047), or parts of the MUN domain (aa 319–901). The first ∼100 aa of Munc13-4 of unknown structure were omitted for simplicity. MHD1 (aa 577–677) and MHD2 (aa 788–894) are two regions within the MUN domain with conserved sequences in all Munc13 proteins. (a) Munc13-4(543-1090), which lacks the C2A and parts of MUN domain but contains the entire region around MHD1/2. (b) Munc13-4(1-899), which lacks the C2B but contains the entire MHD1/2 region. (c) Munc13-4(285-917), which lacks C2A and C2B. (d) Munc13-4(1-780), which is truncated after MHD1 and therefore lacks MHD2-C2B. (e) Munc13-4(781-1090), which lacks the entire N-terminal part including MHD1. Arrowheads mark examples of colocalizations of Munc13-4 and VWF on WPBs. Note that mutants containing the MHD1 and MHD2 regions localize to WPBs, whereas those lacking these sequences do not. Scale bars, 10 µm.