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Fig. 1. Expression of Syt III protein in RBL cells and brain. (A) Crude brain
homogenate (lane 1, 10 µg protein) and whole cell lysates
(1x106 cell equivalents) derived from control (lane 2) or Syt
III-overexpressing (RBL-Syt III+, lane 3) or Syt III-suppressed
(RBL-Syt III-, lane 4) RBL cells were resolved by SDS-PAGE and
immunoblotted using a rabbit polyclonal serum directed against the cytoplasmic
domain of Syt III (1: 1000 dilution). (B) A whole cell lysate
(1x106 cell equivalents) derived from RBL cells transiently
transfected with T7-tagged Syt III cDNA were resolved by SDS-PAGE and
immunoblotted using anti T7 antibodies. (C) Total cell extracts (500 µg)
derived from T7-tagged Syt III expressing cells were subjected to
immunoprecipitation (IP) using anti T7 antibodies. The immune complexes were
resolved by SDS-PAGE and immunoblotted (IB) with either anti T7 antibodies or
anti Syt III antibodies, as indicated. Data represent one of three separate
experiments.