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. 2017 Aug 10;292(39):16199–16210. doi: 10.1074/jbc.M117.807438

Figure 3.

Figure 3.

Association of Stx4 N-terminal peptide reduces endogenous Munc18c binding to Stx4-FL 3xFLAG. A, parental MDA-MB-231 cells, GFP-expressing stable cells, and GFP–Stx4–N-term–expressing stable cells were lysed, and GFP was immunoprecipitated. Immunoprecipitates (IP) were probed for Munc18c (arrow). B+A, beads plus antibody; FLAG IgG, beads plus mouse anti-FLAG antibody plus GFP–Stx4–N-term peptide lysate; Rabbit IgG, rabbit IgG plus Stx4 N-terminal peptide lysate. B, parental and stable cells expressing GFP–Stx4–N-term were transiently transfected with Stx4-FL 3xFLAG for 24 h and lysed, and FLAG was immunoprecipitated. Immunoprecipitates were probed for Munc18c and FLAG (arrow). B+L, beads plus lysate from parental cells. C, densitometry of the amount of Munc18c co-immunoprecipitated relative to Stx4-FL 3xFLAG as shown in B. All data are presented as percent of control ± S.D. The asterisk denotes values significantly different from control (parental cells) (*, p < 0.05). All data represent three or more biological replicates with at least three technical replicates.