Figure 4. Characterization of glycosylated and non-glycosylated forms of Atce1/Tisp40 in mouse testis. (a) Extracts prepared from mouse testis (lane 5), untransfected human kidney 293 cells (lane 1), and 293 cells transiently transfected with plasmids encoding HA-tagged Tisp40ß (lane 2), HA-tagged Tisp40 (lane 3) or untagged Tisp40 (lane 4) were probed with AB1462. The positions of the 42- and 36-kd polypeptides are indicated. (b) Testis extracts (lanes 6–7) and extracts of 293 cells transiently transfected with plasmids encoding untagged forms of Tisp40ß (lanes 2–3) and Tisp40 (lanes 4–5) were incubated in the absence (–) or presence (+) of endoglycosidase H and then analyzed by immunoblotting using AB1462. Untransfected 293 cell extracts are in lane 1. The immunoblots presented here are representative of independent experiments performed using extracts from different mouse testes and independent transfections.