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. Author manuscript; available in PMC: 2014 May 3.
Published in final edited form as: Biochem Biophys Res Commun. 2013 Apr 8;434(2):388–393. doi: 10.1016/j.bbrc.2013.04.001

Figure 4. Palmitoylation of TBC1D3 regulates CUL7-induced TBC1D3 ubiquitination.

Figure 4

HeLa cells were transfected with HA-CUL7, Myc-TBC1D3 full length (FL) or Myc-TBC1D3:C318/325S (double mutant, DM). Cytosol fractions (cyto) were prepared from CUL7-expressing cells and membrane fractions (mem) were prepared from TBC1D3-expressing cells, respectively. The cytosol and membrane fractions were incubated together at room temperature for 45 min. TBC1D3 was immunoprecipitated with anti-Myc antibody. Polyubiquitination of TBC1D3 was analyzed by immunoblot using anti-ubiquitin antibody. The experiment was repeated three times.