Figure 5.

Effects of the T65I mutation on lipid binding. A, erlin complex was immunopurified with anti-HA from αT3 E2KO cell lines stably expressing E2HA^WT or E2HA^T65I, and binding to lipid arrays was assessed by probing with anti-erlin2. The lowest panels show immunoblots of material added to the lipid arrays probed with anti-erlin2 and anti-erlin1, demonstrating that approximately equal amounts of E1HA^WT and E2HA^T65I were used and that erlin1 coimmunoprecipitates with E2HA^WT and E2HA^T65I. B, E2HA^WT and E2HA^T65I were expressed transiently in αT3 E1/E2KO cells and immunopurified with anti-HA, and binding of equal amounts to lipid arrays was assessed with anti-erlin2. The lowest panel shows an immunoblot of material added to the lipid arrays probed with anti-erlin2, demonstrating that approximately equal amounts of E2HA^WT and E2HA^T65I were used.