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

Figure 6.

Figure 6

Starvation promotes MTMR13-dependent RAB21 activation and interaction with VAMP8

  • A RAB21 GTPase activity is transiently activated following starvation. GST:APPL1 used as bait for RAB21-GTP pull-down. Immunoblot of GFP:RAB21 in pull-down (RAB21-GTP) and in cell lysates (RAB21-Tot.).
  • B Starvation-induced RAB21 GTPase activity requires MTMR13, as tested in scramble (−) or MTMR13 siRNA (+)-treated HeLa cells. GST:APPL1 pull-down and immunoblot of activated and total RAB21.
  • C Ratio of activated RAB21-GTP in pull-down versus lysate integrated densities from three independent experiments, normalized to scramble siRNA; SEM.
  • D RAB21 interaction with VAMP8 requires MTMR13, as tested in scramble (−) or MTMR13 siRNA (+)-treated HeLa cells. FLAG IP FLAG:RAB21-WT and immunoblot of co-expressed GFP-tagged VAMP8.
  • E Ratio of VAMP8 in IP versus lysate integrated densities from three independent experiments, normalized to scramble siRNA; SEM. See also Supplementary Fig S5.