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. 1998 May 4;141(3):553–566. doi: 10.1083/jcb.141.3.553

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In vitro and in vivo association of NRP/B with p110^RB. (A) Lysates of SH-SY5Y cells were immunoprecipitated with anti-p110^RB antibody (lanes 2 and 5), analyzed by 7.5% SDS-PAGE, and transferred to Immobilon-P membranes. The membranes were processed for Far Western blotting with the Csk kinase (as a negative control) and NRP/B-purified proteins. Lanes 1 and 4, immunoprecipitations were performed with control antibody; lanes 3 and 6, control Sepharose beads alone. (B) In vivo association of NRP/B with p110^RB: (a) 500 μg of total cell lysates obtained from retinoic acid (10 μM) treated and untreated SH-SY5Y cells were subjected to immunoprecipitation with NRP/B and blotted with monoclonal p110^RB antibody. Two different migrated forms were observed: The slower migrated form (lane 1) represents the hyperphosphorylated p110^RB, and the faster migrated form (lane 2) represents the hypophosphorylated p110^RB. (b) After deprobing the blot in a, the membrane was probed with polyclonal NRP/B antibody.