American Society of Hematology

Fig. 1.

$Fig. 1. Lack of association of Grb2 with the Bcr/Abl Y177F mutant and with v-Abl. / (A) Far Western blot. The indicated Abl proteins were expressed by transient transfection in 293T cells, whole cell extracts fractionated by SDS-PAGE and transferred to nitrocellulose filters, and hybridized with a GST-Grb2 (SH2) fusion protein (left pair) or GST alone (right pair). Bound GST protein was detected by anti-GST antibodies and enhanced chemiluminescence. Filters were subsequently stripped and rehybridized with anti-Abl antibody (right panel of each pair). Molecular weight standards are at left, and the positions of p210 Bcr/Abl and c-Abl proteins are indicated by arrowheads at the right. (B) Co-immunoprecipitation. The indicated Abl proteins were expressed by transient transfection of 293T cells, immunoprecipitated with anti-Grb2 (mock-transfected cells only, left lane) or anti-Abl antibodies (all other samples), fractionated by SDS-PAGE, transferred to nitrocellulose, and hybridized with anti-Abl (top panel) or anti-Grb2 (bottom panel) antibodies. Molecular weight standards are at the right, and the position of Grb2 protein is indicated by the arrowhead.$

Lack of association of Grb2 with the Bcr/Abl Y177F mutant and with v-Abl.

(A) Far Western blot. The indicated Abl proteins were expressed by transient transfection in 293T cells, whole cell extracts fractionated by SDS-PAGE and transferred to nitrocellulose filters, and hybridized with a GST-Grb2 (SH2) fusion protein (left pair) or GST alone (right pair). Bound GST protein was detected by anti-GST antibodies and enhanced chemiluminescence. Filters were subsequently stripped and rehybridized with anti-Abl antibody (right panel of each pair). Molecular weight standards are at left, and the positions of p210 Bcr/Abl and c-Abl proteins are indicated by arrowheads at the right. (B) Co-immunoprecipitation. The indicated Abl proteins were expressed by transient transfection of 293T cells, immunoprecipitated with anti-Grb2 (mock-transfected cells only, left lane) or anti-Abl antibodies (all other samples), fractionated by SDS-PAGE, transferred to nitrocellulose, and hybridized with anti-Abl (top panel) or anti-Grb2 (bottom panel) antibodies. Molecular weight standards are at the right, and the position of Grb2 protein is indicated by the arrowhead.

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