Additional File 2.
Western blot of whole cell lysates of GFP-tagged EHD proteins used in Figure 4. Aliquots of 100 μg of the lysates used for co-immunoprecipitations (co-IP) in Figure 4 were run on the same gel as those in Figure 4, transferred to PVDF membranes, and immunoblotted in parallel with anti-GFP antibodies. (A) Whole cell lysates for Figure 4A. (B) Whole cell lysates for Figure 4B. (C) Control IPs using 1 mg of whole cell lysates (WCL) and co-IPs were carried out as described in Methods using GFP-myotubularian-related protein 3 (MTMR3), Myc-sorting nexin 1 (SNX1), Myc-EHD1 and EHD1-GFP. Lanes 1–3: WCL, 100 μg. Lanes 4–6: 1 mg IP. Relative molecular weight (MW) markers are indicated in kiloDaltons (kD). The heavy chain of the mouse IgG (IgH) is also shown indicating similar levels of antibody (anti-Myc, 9e10) were used for the IP.
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George et al. BMC Cell Biology 2007 8:3 doi:10.1186/1471-2121-8-3