![Fig. 6. MLLC is highly phosphorylated. / (A) MLL is phosphorylated in vivo. BOSC23 cells were transfected with the empty pLNCX vector (vector) or pLNCX MLL(H) [denoted as MLL(H)]. 48 hours after transfection, the cells were metabolically labeled with [32P]-orthophosphate for 3 hours and lysed in lysis buffer. MLLC was immunoprecipitated with the anti-HA antibody 3F10. The precipitates were analyzed by immunoblot with anti-MLL N-terminal antibody (mmN3; left panel) or anti-HA antibody 3F10 (middle panel) or visualized by autoradiography (right panel). Molecular weight markers are on the left. (B) Effects of phosphatase treatment on the mobility of MLLC. BOSC23 cells were transfected with the empty pLNCX vector or pLNCX MLL(H). The cells were lysed in lysis buffer 48 hours after transfection. MLL proteins were immunoprecipitated with the anti-MLL N-terminal antibody (rpN1). Immunoprecipitates were treated with or without calf intestine alkaline phosphatase for 30 minutes and subjected to immunoblotting with the anti-HA antibody 3F10.](https://ash.silverchair-cdn.com/ash/content_public/journal/blood/100/10/10.1182_blood-2002-04-1015/4/h82223465006.jpeg?Expires=1735503905&Signature=ykRhCSSsv-PeMQsTDSOS74fjansD4wLqo2qTZZrFum01-XV35HFYUK7jyrYu6F5lwBlg6pMH5EqQ~Iuduy29wIUWF9qbRS9PLlWCh0AojQ0QwSmaI4dTpthaCVqz-cwm1ZblshgtRXTfs6AeFYZANlWGhn5uj52s8Ca4MdOosHjD~2Uzso8BKxNh1QoqdKAtOmgfxzmZX23WvLtsvGTaPPxXzqoTrsNZlLPYrhJ8kJsODI4NkqkKzGRyrjyR-kP3PDkvZpQPcfW9e1dbpz6550PkGe6-YZEzB835-F0znRv96jBTnuDN92zxkQI4jLwscD346~Obd~LMK~HO0hYL1w__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
MLLC is highly phosphorylated.
(A) MLL is phosphorylated in vivo. BOSC23 cells were transfected with the empty pLNCX vector (vector) or pLNCX MLL(H) [denoted as MLL(H)]. 48 hours after transfection, the cells were metabolically labeled with [32P]-orthophosphate for 3 hours and lysed in lysis buffer. MLLC was immunoprecipitated with the anti-HA antibody 3F10. The precipitates were analyzed by immunoblot with anti-MLL N-terminal antibody (mmN3; left panel) or anti-HA antibody 3F10 (middle panel) or visualized by autoradiography (right panel). Molecular weight markers are on the left. (B) Effects of phosphatase treatment on the mobility of MLLC. BOSC23 cells were transfected with the empty pLNCX vector or pLNCX MLL(H). The cells were lysed in lysis buffer 48 hours after transfection. MLL proteins were immunoprecipitated with the anti-MLL N-terminal antibody (rpN1). Immunoprecipitates were treated with or without calf intestine alkaline phosphatase for 30 minutes and subjected to immunoblotting with the anti-HA antibody 3F10.
