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Fig. 6 | EGFR and Erbb2 bind karyopherins of the classical nuclear-import family. a–e Predicted sensitivity of the systems model to starting protein con- centrations of RanBP3 (a), CAS (b), Ran (c), Imp-α (d), and Imp-β (e). Protein abundances are scaled relative to the concentrations used in the base model (Supplementary Data 4). Steady-state nuclear-to-cytoplasmic (N/C) ratios are shown for 1 µM of the representative cargo described in Fig. 4a. f, g, EGFR inducibly associates with Erbb2 and constitutively associates with KPNA2 and KPNB1. B2B1 cells expressing doxycycline (DOX)-inducible <t>3xFLAG-KPNA2</t> or 3xFLAG- KPNB1 were induced with 1 µg/ml DOX for 24 h and then treated with or without 0.5 µM AP for 1 h before lysis and GluGlu immunoprecipitation of the EGFR chimera. Samples were immunoblotted for the indicated targets (f) and quantified from n = 3 paired biological replicates by densitometry relative to the amount of immuno- precipitated EGFR chimera (g). Differences in overall KPNA2 and KPNB1 co- immunoprecipitation were assessed by two-way ANOVA with replication and AP
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Fig. 6 | EGFR and Erbb2 bind karyopherins of the classical nuclear-import family. a–e Predicted sensitivity of the systems model to starting protein con- centrations of RanBP3 (a), CAS (b), Ran (c), Imp-α (d), and Imp-β (e). Protein abundances are scaled relative to the concentrations used in the base model (Supplementary Data 4). Steady-state nuclear-to-cytoplasmic (N/C) ratios are shown for 1 µM of the representative cargo described in Fig. 4a. f, g, EGFR inducibly associates with Erbb2 and constitutively associates with KPNA2 and KPNB1. B2B1 cells expressing doxycycline (DOX)-inducible <t>3xFLAG-KPNA2</t> or 3xFLAG- KPNB1 were induced with 1 µg/ml DOX for 24 h and then treated with or without 0.5 µM AP for 1 h before lysis and GluGlu immunoprecipitation of the EGFR chimera. Samples were immunoblotted for the indicated targets (f) and quantified from n = 3 paired biological replicates by densitometry relative to the amount of immuno- precipitated EGFR chimera (g). Differences in overall KPNA2 and KPNB1 co- immunoprecipitation were assessed by two-way ANOVA with replication and AP
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Fig. 6 | EGFR and Erbb2 bind karyopherins of the classical nuclear-import family. a–e Predicted sensitivity of the systems model to starting protein con- centrations of RanBP3 (a), CAS (b), Ran (c), Imp-α (d), and Imp-β (e). Protein abundances are scaled relative to the concentrations used in the base model (Supplementary Data 4). Steady-state nuclear-to-cytoplasmic (N/C) ratios are shown for 1 µM of the representative cargo described in Fig. 4a. f, g, EGFR inducibly associates with Erbb2 and constitutively associates with KPNA2 and KPNB1. B2B1 cells expressing doxycycline (DOX)-inducible <t>3xFLAG-KPNA2</t> or 3xFLAG- KPNB1 were induced with 1 µg/ml DOX for 24 h and then treated with or without 0.5 µM AP for 1 h before lysis and GluGlu immunoprecipitation of the EGFR chimera. Samples were immunoblotted for the indicated targets (f) and quantified from n = 3 paired biological replicates by densitometry relative to the amount of immuno- precipitated EGFR chimera (g). Differences in overall KPNA2 and KPNB1 co- immunoprecipitation were assessed by two-way ANOVA with replication and AP
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Fig. 6 | EGFR and Erbb2 bind karyopherins of the classical nuclear-import family. a–e Predicted sensitivity of the systems model to starting protein con- centrations of RanBP3 (a), CAS (b), Ran (c), Imp-α (d), and Imp-β (e). Protein abundances are scaled relative to the concentrations used in the base model (Supplementary Data 4). Steady-state nuclear-to-cytoplasmic (N/C) ratios are shown for 1 µM of the representative cargo described in Fig. 4a. f, g, EGFR inducibly associates with Erbb2 and constitutively associates with KPNA2 and KPNB1. B2B1 cells expressing doxycycline (DOX)-inducible <t>3xFLAG-KPNA2</t> or 3xFLAG- KPNB1 were induced with 1 µg/ml DOX for 24 h and then treated with or without 0.5 µM AP for 1 h before lysis and GluGlu immunoprecipitation of the EGFR chimera. Samples were immunoblotted for the indicated targets (f) and quantified from n = 3 paired biological replicates by densitometry relative to the amount of immuno- precipitated EGFR chimera (g). Differences in overall KPNA2 and KPNB1 co- immunoprecipitation were assessed by two-way ANOVA with replication and AP
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Fig. 6 | EGFR and Erbb2 bind karyopherins of the classical nuclear-import family. a–e Predicted sensitivity of the systems model to starting protein con- centrations of RanBP3 (a), CAS (b), Ran (c), Imp-α (d), and Imp-β (e). Protein abundances are scaled relative to the concentrations used in the base model (Supplementary Data 4). Steady-state nuclear-to-cytoplasmic (N/C) ratios are shown for 1 µM of the representative cargo described in Fig. 4a. f, g, EGFR inducibly associates with Erbb2 and constitutively associates with KPNA2 and KPNB1. B2B1 cells expressing doxycycline (DOX)-inducible 3xFLAG-KPNA2 or 3xFLAG- KPNB1 were induced with 1 µg/ml DOX for 24 h and then treated with or without 0.5 µM AP for 1 h before lysis and GluGlu immunoprecipitation of the EGFR chimera. Samples were immunoblotted for the indicated targets (f) and quantified from n = 3 paired biological replicates by densitometry relative to the amount of immuno- precipitated EGFR chimera (g). Differences in overall KPNA2 and KPNB1 co- immunoprecipitation were assessed by two-way ANOVA with replication and AP

Journal: Nature communications

Article Title: Nucleocytoplasmic transport of active HER2 causes fractional escape from the DCIS-like state.

doi: 10.1038/s41467-023-37914-x

Figure Lengend Snippet: Fig. 6 | EGFR and Erbb2 bind karyopherins of the classical nuclear-import family. a–e Predicted sensitivity of the systems model to starting protein con- centrations of RanBP3 (a), CAS (b), Ran (c), Imp-α (d), and Imp-β (e). Protein abundances are scaled relative to the concentrations used in the base model (Supplementary Data 4). Steady-state nuclear-to-cytoplasmic (N/C) ratios are shown for 1 µM of the representative cargo described in Fig. 4a. f, g, EGFR inducibly associates with Erbb2 and constitutively associates with KPNA2 and KPNB1. B2B1 cells expressing doxycycline (DOX)-inducible 3xFLAG-KPNA2 or 3xFLAG- KPNB1 were induced with 1 µg/ml DOX for 24 h and then treated with or without 0.5 µM AP for 1 h before lysis and GluGlu immunoprecipitation of the EGFR chimera. Samples were immunoblotted for the indicated targets (f) and quantified from n = 3 paired biological replicates by densitometry relative to the amount of immuno- precipitated EGFR chimera (g). Differences in overall KPNA2 and KPNB1 co- immunoprecipitation were assessed by two-way ANOVA with replication and AP

Article Snippet: For KPNA2, a linker was cloned into pEN_TT_miRc2 3xFLAG (Addgene #192305), and KPNA2 into pEN_TT_miRc2 3xFLAG linker.

Techniques: Expressing, Lysis, Immunoprecipitation