EHD protein expression in normal mouse tissues. An 18 week-old male and female C57BL/6 mouse was sacrificed, organs were removed and lysed in tissue lysis buffer as described in Methods. Aliquots of 100 μg tissue lysate were separated using 8% SDS-PAGE and a Western blot was performed using antibodies raised against human EHD proteins. The membrane was serially stripped and reprobed beginning with both EHD1 and EHD4, followed by EHD2, EHD3 and Hsc70 antibodies. The * denotes bands that bled through from the previous blot following stripping. Differential mobility of Hsc70 may represent tissue specific isoforms. Blots shown have exposure times of less than 10 seconds, upon longer exposures, most EHD proteins can be seen in each organ shown. Relative molecular weight (MW) markers are indicated in kiloDaltons (kD). As a loading control, Hsc70 was blotted. M. gland – mammary gland.