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. 2015 Jul 22;156(10):3548–3558. doi: 10.1210/en.2015-1120

Figure 8. Effect of GIP stimulation on GLUT4 translocation with or without Tctex1d2 overexpression and knockdown.

Figure 8.

A, 3T3–L1 adipocytes were untreated or incubated with 0.2 nM insulin for 30 minutes as indicated. In addition, cells were untreated or incubated with 0.2 nM insulin for 30 minutes accompanied with 500 nM GIP for 30 minutes as indicated. Thereafter, the GLUT4 translocation amount was quantitatively estimated as described under Materials and Methods. These data are from 4 independent determinations expressed as the mean ± SD. B, 3T3–L1 adipocytes were transfected with either the empty vector (pcDNA3.1) or pcDNA3.1-Tctex1d2 as described under Materials and Methods. Cells were either untreated or incubated with 100 nM insulin for 30 minutes as indicated. The other set of cells were either untreated or incubated with 0.2 nM insulin for 30 minutes as indicated and incubated with 500 nM GIP for 30 minutes as indicated. Thereafter, the GLUT4 translocation amount was quantitatively estimated as described under Materials and Methods. These data are from 7 independent determinations expressed as the mean ± SD. C and D, scrambled (lanes 3 and 6) or Tctex1d2-specific RNAi (lanes 1, 2, 4, and 5)–transfected 3T3–L1 adipocytes were either left untreated (lanes 1 and 4) or incubated with 0.2 nM insulin for 30 minutes (lane 2 and 5) or 0.2 nM insulin for 30 minutes accompanied with 500 nM GIP for 30 minutes (lanes 3 and 6) as indicated. Cell extracts were prepared and immunoblotted for Doc2b, syntaxin 4, and Tctex1d2. In parallel, syntaxin 4 was immunoprecipitated, and the resultant precipitates were immunoblotted for Doc2b, syntaxin 4, and Tctex1d2. These are representative immunoblots performed independently 3 times. H.C., heavy chain; A.U., arbitrary units.