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. Author manuscript; available in PMC: 2017 Sep 1.
Published in final edited form as: Traffic. 2016 Jul 11;17(9):1027–1041. doi: 10.1111/tra.12418

Figure 4. Dominant negative (S25N) Rab11 inhibits stimulated exocytosis monitored by VAMP8-pHl or β-Hex-pHl, which can be rescued by cytochalasin D (cytoD).

Figure 4

(A) RBL-2H3 cells were co-transfected with VAMP8-pHluorin and wild-type (WT) or dominant negative (S25N) mCh-Rab11, and sensitized with anti-DNP IgE (for stimulation with Ag) before harvest. Cells were treated with 2 μM cytoD or vehicle (DMSO, -cytoD) for 5 minutes before exocytosis was stimulated with 100 ng/ml DNP-BSA (Ag). Fluorescence was monitored spectroscopically. Data were normalized as described in Materials and Methods. Bar graph shows integrated fluorescence values expressed as a ratio (S25N/WT) to quantify rescue of S25N Rab11-mediated inhibition by cytoD. (B) Cells were prepared and analyzed as in (A) except that 1 μM A23187 (A23) was added in place of Ag. (C) Cells were prepared and analyzed as in (A) except that β-Hex-pHl was transfected and monitored instead of VAMP8-pHl. In all cases: the average of three experiments at t=300 s is shown at right, error bars are standard deviations, and p-values were calculated using the unpaired t-test.