Figure 5. Myo5B is required for Slp4a–Stx3 interaction. (A) Interaction between v-SNARE Vamp3, Rab11a, Rab8a, and Rab3b is not affected by Myo5B depletion. (B) Interaction between v-SNARE–like Slp4a and t-SNARE Stx3 is abolished upon Myo5B depletion. The interaction with Rab8a and Rab3b is not affected. (C) v-SNARE Stx3 fails to pull down proteins of the apical cargo trafficking pathway, Sec15, Rab11a, Rab8a, or Rab3b, when Myo5B is deleted. Interaction with t-SNARE Vamp3 remains unaffected (compare with A). (D) Interaction between SM protein Munc18-2 with Stx3 and apical v-SNARE Vamp7 with Stx3 is strongly reduced upon deletion of Myo5B, whereas Vamp8/Stx3 and Vamp3/Stx3 interaction seems unaffected. (E) Active Rab11a affinity purifies Vamp7 but active Rab8a or Myo5B. Munc18-2 and Vamp8 do not interact with any of the bait proteins. (A–E) Strep-Tactin was used for coaffinity purification (co-AP).
Figure 5.

Myo5B is required for Slp4a–Stx3 interaction. (A) Interaction between v-SNARE Vamp3, Rab11a, Rab8a, and Rab3b is not affected by Myo5B depletion. (B) Interaction between v-SNARE–like Slp4a and t-SNARE Stx3 is abolished upon Myo5B depletion. The interaction with Rab8a and Rab3b is not affected. (C) v-SNARE Stx3 fails to pull down proteins of the apical cargo trafficking pathway, Sec15, Rab11a, Rab8a, or Rab3b, when Myo5B is deleted. Interaction with t-SNARE Vamp3 remains unaffected (compare with A). (D) Interaction between SM protein Munc18-2 with Stx3 and apical v-SNARE Vamp7 with Stx3 is strongly reduced upon deletion of Myo5B, whereas Vamp8/Stx3 and Vamp3/Stx3 interaction seems unaffected. (E) Active Rab11a affinity purifies Vamp7 but active Rab8a or Myo5B. Munc18-2 and Vamp8 do not interact with any of the bait proteins. (A–E) Strep-Tactin was used for coaffinity purification (co-AP).

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