American Society of Hematology

Fig. 2.

$Fig. 2. Generation of the stable cell lines expressing membrane-associated forms of Syk. / (A) The Syk-negative variant of the RBL-2H3 cells (B2 cells) was stably transfected with the various Syk cDNAs shown in Figure 1 and selected with G418. Two positive cloned lines expressing each of the different forms of Syk were selected for further analysis. Cell lysates from RBL-2H3, parental B2, and selected cloned lines were immunoblotted with anti-Syk and anti-FcεRIβ antibodies. Molecular size markers are indicated at the left in kilodaltons. (B) Cell lysates from RBL-2H3 cells and the transfected cell lines expressing wild-type Syk (clone 5), myr-Syk (A28), and myr-SykΔS (E15) were immunoprecipitated (IP) with either anti-Syk or anti-Src (LA074) antibody. Immunoprecipitated proteins were analyzed by immunoblotting with anti-Syk antibody. (C) Membrane (M) and cytosolic (C) fractions of RBL-2H3 cells and the transfected cell lines were analyzed by immunoblotting with anti-Syk and anti-FcεRIβ antibodies.$

Generation of the stable cell lines expressing membrane-associated forms of Syk.

(A) The Syk-negative variant of the RBL-2H3 cells (B2 cells) was stably transfected with the various Syk cDNAs shown in Figure 1 and selected with G418. Two positive cloned lines expressing each of the different forms of Syk were selected for further analysis. Cell lysates from RBL-2H3, parental B2, and selected cloned lines were immunoblotted with anti-Syk and anti-FcεRIβ antibodies. Molecular size markers are indicated at the left in kilodaltons. (B) Cell lysates from RBL-2H3 cells and the transfected cell lines expressing wild-type Syk (clone 5), myr-Syk (A28), and myr-SykΔS (E15) were immunoprecipitated (IP) with either anti-Syk or anti-Src (LA074) antibody. Immunoprecipitated proteins were analyzed by immunoblotting with anti-Syk antibody. (C) Membrane (M) and cytosolic (C) fractions of RBL-2H3 cells and the transfected cell lines were analyzed by immunoblotting with anti-Syk and anti-FcεRIβ antibodies.

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