(A) Immunoblots of RAPSYN and BCR-ABL in the peripheral blood mononuclear cells (PBMCs) of clinical samples. (B) Immunoblots of RAPSYN and BCR-ABL in Ph+ leukemic cells and normal bone marrow …
Original file for the Western blot analysis in Figure 1A (anti-BCR-ABL, anti-RAPSYN, anti-GAPDH).
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Original file for the Western blot analysis in Figure 1B (anti-BCR-ABL, anti-RAPSYN, anti-β-Tubulin).
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Original file for the Western blot analysis in Figure 1I (anti-BCR-ABL, anti-RAPSYN, anti-β-Tubulin).
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Original file for the Western blot analysis in Figure 1J (anti-BCR-ABL, anti-RAPSYN, anti-β-Tubulin).
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(A) Analyses of RAPSN mRNA levels in peripheral blood mononuclear cells (PBMCs) of healthy donors and non-leukemic patients compared to those of patients with chronic myeloid leukemia (CML) from …
Original file for the Western blot analysis in Figure 1—figure supplement 1E (anti-RAPSYN, anti-GAPDH).
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(A) Co-immunoprecipitation of BCR-ABL and RAPSYN in leukemic cells. (B) Immunoblots of GST and His after immunoprecipitation of His or GST in HEK293T cells transfected with His-tagged BCR-ABL and …
Original file for the Western blot analysis in Figure 2A.
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Original file for the Western blot analysis in Figure 2B.
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Original file for the Western blot analysis in Figure 2C.
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Original file for the Western blot analysis in Figure 2D.
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Original file for the Western blot analysis in Figure 2E.
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Original file for the Western blot analysis in Figure 2F.
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Original file for the Western blot analysis in Figure 2G.
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Original file for the Western blot analysis in Figure 2H.
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Original file for the Western blot analysis in Figure 2I.
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Original file for the Western blot analysis in Figure 2J.
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Original file for the Western blot analysis in Figure 2K.
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Original file for the Western blot analysis in Figure 2L.
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Original file for the Western blot analysis in Figure 2M.
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Original file for the Western blot analysis in Figure 2N.
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(A) Immunoblots of AChR subunits α7, M2, M3, and M4 in Ph+ leukemia cells (K562, KU812, and MEG-01) compared to normal bone marrow stromal cells (HS-5). (B) Immunoblotting analyses of AChR subunit α7…
Original file for the Western blot analysis in Figure 2—figure supplement 1A.
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Original file for the Western blot analysis in Figure 2—figure supplement 1B.
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Original file for the Western blot analysis in Figure 2—figure supplement 1D.
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Original file for the Western blot analysis in Figure 2—figure supplement 1E.
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Original file for the Western blot analysis in Figure 2—figure supplement 1F.
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Original file for the Western blot analysis in Figure 2—figure supplement 1G.
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Original file for the Western blot analysis in Figure 2—figure supplement 1H.
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Original file for the Western blot analysis in Figure 2—figure supplement 1I.
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The neddylation at K257, K500, K739, K802, K1025, K1135, K1590, and K1990 is, respectively, presented with the numbering at Lys residue. Detected peptide sequences are indicated in blue (b ions) and …
(A) Immunoblots of BCR-ABL in leukemic cells treated with MLN4924 or DMSO for 24 hr and corresponding quantification of three independent replicates. (B) Immunoblots of BCR-ABL in K562 WT and RAPSN …
Original file for the Western blot analysis in Figure 3A.
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Original file for the Western blot analysis in Figure 3B.
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Original file for the Western blot analysis in Figure 3C.
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Original file for the Western blot analysis in Figure 3D.
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Original file for the Western blot analysis in Figure 3E.
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Original file for the Western blot analysis in Figure 3F.
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Original file for the Western blot analysis in Figure 3G.
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Original file for the Western blot analysis in Figure 3H.
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Original file for the Western blot analysis in Figure 3I.
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Immunoblotting analyses of BCR-ABL in KU812 and Jurkat cells treated with MLN4924 or DMSO for 24 hr (n=3). Error bars, mean ± standard deviation (SD); *p < 0.05; ***p < 0.001; Student’s t-test. See …
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(A) Assessment of RAPSYN phosphorylation levels in leukemic cells treated with saracatinib or DMSO for 24 hr. (B) Assessment of RAPSYN phosphorylation levels in leukemic cells transduced with shSRC …
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Original file for the Western blot analysis in Figure 4C.
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Original file for the Western blot analysis in Figure 4D.
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Original file for the Western blot analysis in Figure 4G.
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Original file for the Western blot analysis in Figure 4K.
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(A) Assessment of RAPSYN phosphorylation in KU812 cells treated with saracatinib or DMSO for 24 hr. (B) Liquid chromatography–mass spectrometry (LC–MS/MS) spectra of trypsin-digested RAPSYN …
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(A) Immunoblots of BCR-ABL neddylation levels in leukemic cells treated with saracatinib or DMSO for 24 hr. (B) Immunoblots of BCR-ABL neddylation levels in leukemic cells transduced with shSRC or …
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Original file for the Western blot analysis in Figure 5F.
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(A) Cytotoxicity induced by shSRC #2-mediated SRC knockdown in leukemic cells. (B) Rescue of leukemic cells from shSRC #2-induced toxicity by exogenous expression of SRC cDNA. (C) Rescue of leukemic …
Reagent type (species) or resource | Designation | Source or reference | Identifiers | Additional information |
---|---|---|---|---|
Strain, strain background (M. musculus) | Mouse: NOD/ShiLtJGpt-Prkdcem26Cd52Il2rgem26Cd22/Gpt | GemPharmatech | Cat# CB101 | |
Cell line (Homo sapiens) | Human bone marrow stromal cell HS-5 (male) | ATCC | Cat# CRL11882; RRID:CVCL_3720 | |
Cell line (Homo sapiens) | Human chronic myelogenous leukemia K562 (female) | COBIOER | Cat# CBP60529 | |
Cell line (Homo sapiens) | Human chronic myelogenous leukemia MEG-01 (male) | COBIOER | Cat# CBP61104 | |
Cell line (Homo sapiens) | Human chronic myelogenous leukemia KU812 (male) | COBIOER | Cat# CBP60732 | |
Cell line (Homo sapiens) | HEK-293T | KeyGEN BioTECH | Cat# KG405 | |
Antibody | Mouse monoclonal anti-RAPSYN (clone 1234) | Abcam | Cat# ab11423; RRID:AB_298028 | 1:1000 |
Antibody | Rabbit polyclonal anti-RAPSYN (clone 118491) | Abcam | Cat# ab118491; RRID:AB_10899872 | 1:1000 |
Antibody | Mouse monoclonal anti-6X His tag (clone HIS.H8) | Abcam | Cat# ab18184; RRID:AB_444306 | 1:1000 |
Antibody | Mouse monoclonal anti-BCR-ABL (clone 7C6) | Abcam | Cat# ab187831 | 1:1000 |
Antibody | Rabbit monoclonal anti-SRC Family (phosphoY418) | Abcam | Cat# ab40660 | 1:1000 |
Antibody | Rabbit monoclonal anti-NEDD8 (clone 19E3) | Cell Signaling Technology | Cat# 2754; RRID:AB_659972 | 1:1000 |
Antibody | Rabbit monoclonal anti-HA-Tag (clone C29F4) | Cell Signaling Technology | Cat# 3724; RRID:AB_1549585 | 1:1000 |
Antibody | Rabbit monoclonal anti-GFP (clone D5.1) | Cell Signaling Technology | Cat# 2956; RRID:AB_1196615 | 1:1000 |
Antibody | Rabbit monoclonal anti-GST (clone 91G1) | Cell Signaling Technology | Cat#2625 | 1:1000 |
Antibody | Mouse monoclonal anti-Myc-Tag (clone 9B11) | Cell Signaling Technology | Cat#2276; RRID:AB_331783 | 1:1000 |
Antibody | Rabbit monoclonal anti-SRC (clone 36D10) | Cell Signaling Technology | Cat# 2109; RRID:AB_2106059 | 1:1000 |
Antibody | Rabbit monoclonal anti-Flag (DYKDDDDK) Tag (clone D6W5B) | Cell Signaling Technology | Cat# 14793; RRID:AB_2572291 | 1:1000 |
Antibody | Rabbit monoclonal anti-GAPDH (clone 14C10) | Cell Signaling Technology | Cat# 2128; RRID:AB_823664 | 1:2000 |
Antibody | Anti-mouse IgG, HRP-linked antibody | Cell Signaling Technology | Cat# 7076 S; RRID:AB_330924 | 1:5000 |
Antibody | Anti-rabbit IgG, HRP-linked antibody | Cell Signaling Technology | Cat# 7074 S; RRID:AB_2099233 | 1:5000 |
Antibody | Normal Mouse IgG | Santa Cruz Biotechnology | Cat# sc-2025; RRID:AB_737182 | 1:100 |
Antibody | Mouse monoclonal anti-c-CBL (clone A-9) | Santa Cruz Biotechnology | Cat# SC-1651; RRID:AB_2244054 | 1:1000 |
Antibody | Mouse monoclonal anti-Phosphotyrosine Antibody (clone 4G10) | Sigma-Aldrich | Cat# 05-321 X; RRID:AB_568858 | 1:1000 |
Antibody | Rabbit monoclonal anti-AChRα7 | Santa Cruz Biotechnology | Cat# SC-58607; RRID:AB_784835 | 1:1000 |
Antibody | Rabbit monoclonal anti-mAChR M2 | Santa Cruz Biotechnology | Cat# SC-33712; RRID:AB_673789 | 1:1000 |
Antibody | Mouse monoclonal anti-mAChR M3 | Santa Cruz Biotechnology | Cat# SC-518107 | 1:1000 |
Antibody | Rbbit polyclonal anti-mAChR M4 | HUABIO | Cat# ER1906-24; | 1:1000 |
Strain, strain background (Escherichia coli) | DH5-alpha | TIANGEN | Cat# CB101 | |
Strain, strain background (Escherichia coli) | ArcticExpress (DE3) pRARE2 | ANGYUBIO | Cat# AYBIO-G6023 | |
Transfected construct (human) | Plasmid: pcDNA 3.1(+) mammalian expression vector | Invitrogen | Cat# V79020 | |
Strain, strain background (human) | Plasmid: pd1-EGFP-N1 mammalian expression vector | Clontech | Cat# 6073-1 | |
Transfected construct (Escherichia coli) | pGEX-4T-1 bacterial expression vector | Addgene | 27-4580-01 |
Sequences of shRNA and primer pairs used for real-time PCR to determine the mRNA levels.