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. 2015 Feb 3;16(3):297–311. doi: 10.15252/embr.201439464

Figure 5.

Figure 5

Starvation promotes RAB21 and Sbf/MTMR13 interactions with dVamp7/VAMP8

  • A RAB21 interacts with VAMP8 and VAMP7 in HeLa cells. RAB21 interaction with VAMP8 is increased by starvation. FLAG IP FLAG:Rab21-WT and immunoblot of co-expressed GFP-tagged VAMP8 or VAMP7.
  • B VAMP8 interacts preferentially with active RAB21 forms in response to starvation (90 min). FLAG IP FLAG:RAB21-WT, RAB21-CA or RAB21-DN and immunoblot of co-expressed GFP-tagged VAMP8.
  • C MTMR13 interacts with VAMP8 under starvation (60 min). MTMR13 IP of Myc:MTMR13 and immunoblot of co-expressed GFP:VAMP8.
  • D MTMR13-DENN interacts preferentially with inactive RAB21 in response to starvation (90 min). FLAG IP FLAG:RAB21-WT, RAB21-CA or RAB21-DN and immunoblot of co-expressed GFP:MTMR13-DENN (GEF) domain.
  • E-G Protein interactions in co-IP from fed or starved Drosophila third instar larvae. Starvation induces in vivo interaction between (E) endogenous Drosophila Rab21 (IP) and Sbf, (F) endogenous Rab21 (IP) and fat body-expressed GFP:Vamp7 (anti-GFP IB) and (G) endogenous Sbf (IP) and fat body-expressed GFP:VAMP7 (anti-GFP IB). See also Supplementary Fig S5.