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. 2017 Nov 17;37(2):300–320. doi: 10.15252/embj.201796484

Figure EV1. In vitro kinase assays and Syntaxin binding of Munc18‐1 variants.

Figure EV1

  1. In vitro kinase assay in which lysates from HEK cells expressing n‐Src or Munc18‐1 were combined and incubated in the presence of PTP inhibitor vanadate at 4°C. Samples were denatured after different incubation times and Munc18‐1 was immunoprecipitated and immunoblotted against phosphotyrosine.
  2. Same as (A) but using neuronal Munc18‐1 from brain lysate and incubation at room temperature.
  3. Y473 mutants all bind Syntaxin1. Munc18‐1 variants were co‐expressed with Syntaxin1 in HEK293T cells. Munc18‐1 and Munc18‐1‐bound proteins were immunoprecipitated from cell lysate and immunoblotted for detection.

Source data are available online for this figure.