Fig. 1 | Leukemia

Fig. 1

From: The phosphatase UBASH3B/Sts-1 is a negative regulator of Bcr-Abl kinase activity and leukemogenesis

Fig. 1

The Sts-1 phosphatase interacts with and dephosphorylates Bcr-Abl. a SupB15 was treated with imatinib (2 µM for 6 h). ABL1 immunoprecipitates (left panel) from total cell lysates and 5% input fraction (right panel) were immunoblotted with the indicated antibodies. b Ba/F3 cells transduced with STS-1 were additionally transduced with BCR-ABL1 and treated with imatinib (1 µM for 6 h). STS-1 immunoprecipitates (right panel) from total cell lysates and 5% input fraction (left panel) were immunoblotted with the indicated antibodies. c Ba/F3 cells transduced with BCR-ABL1 were additionally transduced with STS-1 and treated with imatinib (1 µM for 6 h). ABL1 immunoprecipitates (right panel) from total cell lysates and 5% input fraction (left panel) were immunoblotted with the indicated antibodies. d Ba/F3 cells transduced with full-length Bcr-Abl, the Abl-portion (#ABL1) only, the Bcr oligomerization domain directly fused to the Abl-portion (BCC-ABL1) or only the Bcr-portion (BCR) was additionally transduced with STS-1. ABL1 (upper panels) and STS-1 (lower panels) immunoprecipitates from total cell lysates were immunoblotted with the indicated antibodies. e HEK293 cells were cotransfected with BCR-ABL1 wt, Bcr-Abl kinase-dead (D382N; BCR-ABLΔK) or BCR-ABL1 lacking the C-terminal last exon region (BCR-ABLΔLE) with STS-1 wt and point mutations in functional domains (UBA (W72E), SH3 (W295A), and PGM (H391A)). ABL1 immunoprecipitates were analyzed by immunoblotting using the indicated antibodies. f Ba/F3 cells retrovirally expressing BCR-ABL1 were transfected with STS-1 or empty vector. Whole-cell lysates were then analyzed with the indicated antibodies. g HEK293 cells were cotransfected with BCR-ABL1 wt or an oligomerization-defective BCR-ABL1 lacking the coiled-coil domain (ΔCC-BCR-ABL1) with either STS-1 wt or a phosphatase-dead STS-1 mutant (H391A). Total cell lysates were analyzed by immunoblotting using the indicated antibodies

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