Figure 9From: Shared as well as distinct roles of EHD proteins revealed by biochemical and functional comparisons in mammalian cells and C. elegansDifferential effects of wild type and ΔEH mutants of EHD1, 3, & 4 versus EHD2 on transferrin exit from the ERC. Untransfected HeLa cells or cells transiently transfected with Myc-EHD1 or Myc-EHD1 ΔEH were loaded with Transferrin-coupled Alexa Fluor 488 (Tf, green) in internalization buffer at 37°C for 30 minutes 24 h after transfection, washed with ice-cold PBS and chased with serum-containing medium. The cells were fixed at various time points, stained with antibodies for Myc (9E10, red) and scanned on a confocal microscope. Similar experiments with EHD2-4 and EHD2-4 ΔEH are shown in Additional File 5. (B) Cells expressing Myc-EHD proteins (black bars) or Myc-EHD ΔEH proteins (grey bars) from a representative experiment were counted with respect to Tf retention after 60 min of chase. At least 200 cells were counted in each case.Back to article page