JCS007310 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1AB
• Supplemental Figure S1CD -

  Fig. S1. Overexpression of Rip11 inhibits insulin-stimulated translocation of GLUT4 to, and fusion with, the plasma membrane. 3T3-L1 adipocytes were electroporated with plasmids encoding HA-GLUT4-GFP in the presence of either mRFP-Rip11 or mRFP vector, as indicated. 24 hours later, the cells were treated in the absence or presence of 100 nM insulin for 30 minutes. The cells were fixed and stained with an antibody against HA to detect surface-localised GLUT4. The figure shows confocal images of the distribution of GFP fluorescence (green) in insulin-stimulated cells also stained with antibodies against HA (red).

• Supplemental Figure S2 -

  Fig. S2. Knockdown of Rip11 has no effect on the expression or subcellular distribution of endogenous GLUT4. 3T3-L1 adipocytes were electroporated with a scrambled siRNA duplex or with either Rip11A or Rip11B duplexes targeted at different regions of the mRNA encoding Rip11. 24 hours later, the cells were fixed and stained for the presence of endogenous GLUT4 using an antibody raised against the C-terminus of the protein (a gift of Geoff Holman, University of Bath, UK). The figure shows images of the cells taken at equivalent magnifications and gain/offset.