brg1 antibody Abcam Search Results


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  • 92
    Abcam antibody against brg1
    The PTEN/AKT/GSK3β axis modulates <t>BRG1</t> stability through the FBXW7-dependent ubiquitin proteasome pathway. ( A ) Sequence alignment of the putative GSK3β phosphorylation sites at S1417 and S1421 of BRG1. ( B ) In vitro kinase assays depicting major GSK3β phosphorylation sites in BRG1. ( C ) IB analysis of WCL and immunoprecipitates from control and PTEN-overexpressing PC3 cells treated with CHIR-99021 or λ-phosphatase as indicated. ( D ) Lysates from control and FBXW7-overexpressing cells were subjected to IP with an anti-BRG1 antibody, and ubiquitinated BRG1 was detected by an anti-Ub antibody. ( E ) IB analysis of PC3 cells transfected with scramble or AKT oligonucleotides with or without FBXW7 KD (shFBXW7). ( F ) Flag-tagged WT, BRG1-SA, and BRG1-SD proteins were incubated with SCF-FBXW7 complex as indicated and then subjected to Western blotting. ( G ) IB analysis of the indicated protein in WCL and immunoprecipitates from 293T cells transfected with HA-tagged FBXW7 and Flag-tagged BRG1, BRG-SA, or BRG1-SD. ( H ) IB analysis of the indicated protein in WT, SA, and SD cells. ( I ) WT, SA, and SD cell lysates were subjected to IP with the indicated antibodies. ( J ) Volume of subcutaneous tumors derived from WT, SA, and SD cells ( n = 6, 2-way ANOVA followed by Tukey’s multiple comparisons test). Scale bar: 1 cm. ( K ) Representative image of BRG1, P-1417/1421, and PTEN expression in lysates from PCa samples (upper panel). Pearson’s correlations among BRG1, P-1417/1421, and PTEN in PCa specimens are summarized in the heatmap ( n = 30). ** P
    Antibody Against Brg1, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    85
    Abcam smarca4 brg1
    The PTEN/AKT/GSK3β axis modulates <t>BRG1</t> stability through the FBXW7-dependent ubiquitin proteasome pathway. ( A ) Sequence alignment of the putative GSK3β phosphorylation sites at S1417 and S1421 of BRG1. ( B ) In vitro kinase assays depicting major GSK3β phosphorylation sites in BRG1. ( C ) IB analysis of WCL and immunoprecipitates from control and PTEN-overexpressing PC3 cells treated with CHIR-99021 or λ-phosphatase as indicated. ( D ) Lysates from control and FBXW7-overexpressing cells were subjected to IP with an anti-BRG1 antibody, and ubiquitinated BRG1 was detected by an anti-Ub antibody. ( E ) IB analysis of PC3 cells transfected with scramble or AKT oligonucleotides with or without FBXW7 KD (shFBXW7). ( F ) Flag-tagged WT, BRG1-SA, and BRG1-SD proteins were incubated with SCF-FBXW7 complex as indicated and then subjected to Western blotting. ( G ) IB analysis of the indicated protein in WCL and immunoprecipitates from 293T cells transfected with HA-tagged FBXW7 and Flag-tagged BRG1, BRG-SA, or BRG1-SD. ( H ) IB analysis of the indicated protein in WT, SA, and SD cells. ( I ) WT, SA, and SD cell lysates were subjected to IP with the indicated antibodies. ( J ) Volume of subcutaneous tumors derived from WT, SA, and SD cells ( n = 6, 2-way ANOVA followed by Tukey’s multiple comparisons test). Scale bar: 1 cm. ( K ) Representative image of BRG1, P-1417/1421, and PTEN expression in lysates from PCa samples (upper panel). Pearson’s correlations among BRG1, P-1417/1421, and PTEN in PCa specimens are summarized in the heatmap ( n = 30). ** P
    Smarca4 Brg1, supplied by Abcam, used in various techniques. Bioz Stars score: 85/100, based on 12 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    80
    Abcam anti smarca4 brg1 n terminus
    The PTEN/AKT/GSK3β axis modulates <t>BRG1</t> stability through the FBXW7-dependent ubiquitin proteasome pathway. ( A ) Sequence alignment of the putative GSK3β phosphorylation sites at S1417 and S1421 of BRG1. ( B ) In vitro kinase assays depicting major GSK3β phosphorylation sites in BRG1. ( C ) IB analysis of WCL and immunoprecipitates from control and PTEN-overexpressing PC3 cells treated with CHIR-99021 or λ-phosphatase as indicated. ( D ) Lysates from control and FBXW7-overexpressing cells were subjected to IP with an anti-BRG1 antibody, and ubiquitinated BRG1 was detected by an anti-Ub antibody. ( E ) IB analysis of PC3 cells transfected with scramble or AKT oligonucleotides with or without FBXW7 KD (shFBXW7). ( F ) Flag-tagged WT, BRG1-SA, and BRG1-SD proteins were incubated with SCF-FBXW7 complex as indicated and then subjected to Western blotting. ( G ) IB analysis of the indicated protein in WCL and immunoprecipitates from 293T cells transfected with HA-tagged FBXW7 and Flag-tagged BRG1, BRG-SA, or BRG1-SD. ( H ) IB analysis of the indicated protein in WT, SA, and SD cells. ( I ) WT, SA, and SD cell lysates were subjected to IP with the indicated antibodies. ( J ) Volume of subcutaneous tumors derived from WT, SA, and SD cells ( n = 6, 2-way ANOVA followed by Tukey’s multiple comparisons test). Scale bar: 1 cm. ( K ) Representative image of BRG1, P-1417/1421, and PTEN expression in lysates from PCa samples (upper panel). Pearson’s correlations among BRG1, P-1417/1421, and PTEN in PCa specimens are summarized in the heatmap ( n = 30). ** P
    Anti Smarca4 Brg1 N Terminus, supplied by Abcam, used in various techniques. Bioz Stars score: 80/100, based on 5 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    97
    Santa Cruz Biotechnology smarca4
    The PTEN/AKT/GSK3β axis modulates <t>BRG1</t> stability through the FBXW7-dependent ubiquitin proteasome pathway. ( A ) Sequence alignment of the putative GSK3β phosphorylation sites at S1417 and S1421 of BRG1. ( B ) In vitro kinase assays depicting major GSK3β phosphorylation sites in BRG1. ( C ) IB analysis of WCL and immunoprecipitates from control and PTEN-overexpressing PC3 cells treated with CHIR-99021 or λ-phosphatase as indicated. ( D ) Lysates from control and FBXW7-overexpressing cells were subjected to IP with an anti-BRG1 antibody, and ubiquitinated BRG1 was detected by an anti-Ub antibody. ( E ) IB analysis of PC3 cells transfected with scramble or AKT oligonucleotides with or without FBXW7 KD (shFBXW7). ( F ) Flag-tagged WT, BRG1-SA, and BRG1-SD proteins were incubated with SCF-FBXW7 complex as indicated and then subjected to Western blotting. ( G ) IB analysis of the indicated protein in WCL and immunoprecipitates from 293T cells transfected with HA-tagged FBXW7 and Flag-tagged BRG1, BRG-SA, or BRG1-SD. ( H ) IB analysis of the indicated protein in WT, SA, and SD cells. ( I ) WT, SA, and SD cell lysates were subjected to IP with the indicated antibodies. ( J ) Volume of subcutaneous tumors derived from WT, SA, and SD cells ( n = 6, 2-way ANOVA followed by Tukey’s multiple comparisons test). Scale bar: 1 cm. ( K ) Representative image of BRG1, P-1417/1421, and PTEN expression in lysates from PCa samples (upper panel). Pearson’s correlations among BRG1, P-1417/1421, and PTEN in PCa specimens are summarized in the heatmap ( n = 30). ** P
    Smarca4, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 97/100, based on 94 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    87
    Abcam brg1 biotin
    The PTEN/AKT/GSK3β axis modulates <t>BRG1</t> stability through the FBXW7-dependent ubiquitin proteasome pathway. ( A ) Sequence alignment of the putative GSK3β phosphorylation sites at S1417 and S1421 of BRG1. ( B ) In vitro kinase assays depicting major GSK3β phosphorylation sites in BRG1. ( C ) IB analysis of WCL and immunoprecipitates from control and PTEN-overexpressing PC3 cells treated with CHIR-99021 or λ-phosphatase as indicated. ( D ) Lysates from control and FBXW7-overexpressing cells were subjected to IP with an anti-BRG1 antibody, and ubiquitinated BRG1 was detected by an anti-Ub antibody. ( E ) IB analysis of PC3 cells transfected with scramble or AKT oligonucleotides with or without FBXW7 KD (shFBXW7). ( F ) Flag-tagged WT, BRG1-SA, and BRG1-SD proteins were incubated with SCF-FBXW7 complex as indicated and then subjected to Western blotting. ( G ) IB analysis of the indicated protein in WCL and immunoprecipitates from 293T cells transfected with HA-tagged FBXW7 and Flag-tagged BRG1, BRG-SA, or BRG1-SD. ( H ) IB analysis of the indicated protein in WT, SA, and SD cells. ( I ) WT, SA, and SD cell lysates were subjected to IP with the indicated antibodies. ( J ) Volume of subcutaneous tumors derived from WT, SA, and SD cells ( n = 6, 2-way ANOVA followed by Tukey’s multiple comparisons test). Scale bar: 1 cm. ( K ) Representative image of BRG1, P-1417/1421, and PTEN expression in lysates from PCa samples (upper panel). Pearson’s correlations among BRG1, P-1417/1421, and PTEN in PCa specimens are summarized in the heatmap ( n = 30). ** P
    Brg1 Biotin, supplied by Abcam, used in various techniques. Bioz Stars score: 87/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    82
    Abcam immunostaining anti brg1
    <t>Brg1</t> expression in cochlea and conditional Brg1 inactivation in cochlear HCs. ( A ) Transverse sections of P4 cochlea stained with Brg1 (green), the OHC marker Prestin (red) and DAPI (blue, nuclei). Brg1 was specifically deleted in Atoh1-Brg1 −/− HCs. Inserts represent higher magnification views of the boxed areas. Brackets indicate OHCs, and arrowheads indicate <t>IHC.</t> Abbreviations: RM, Reissner’s membrane; SV, stria vascularis; OC, organ of Corti; SG, spiral ganglia. Scale bars: 100 μm. ( B ) Efficiency of Brg1 deletion shown by whole mount cochlea of P1 and P4 mice stained with Brg1 (green), the HC marker Myosin7a (red) and DAPI (blue, nuclei). Brg1 was sill detected in P1 Atoh1-Brg1 −/− HCs with a lower signal than in control HCs while in P4 Atoh1-Brg1 −/− HCs, Brg1 was absent in all HCs. Scale bars: 20 μm.
    Immunostaining Anti Brg1, supplied by Abcam, used in various techniques. Bioz Stars score: 82/100, based on 7 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    92
    Abcam rabbit anti brg1
    <t>Brg1</t> expression in cochlea and conditional Brg1 inactivation in cochlear HCs. ( A ) Transverse sections of P4 cochlea stained with Brg1 (green), the OHC marker Prestin (red) and DAPI (blue, nuclei). Brg1 was specifically deleted in Atoh1-Brg1 −/− HCs. Inserts represent higher magnification views of the boxed areas. Brackets indicate OHCs, and arrowheads indicate <t>IHC.</t> Abbreviations: RM, Reissner’s membrane; SV, stria vascularis; OC, organ of Corti; SG, spiral ganglia. Scale bars: 100 μm. ( B ) Efficiency of Brg1 deletion shown by whole mount cochlea of P1 and P4 mice stained with Brg1 (green), the HC marker Myosin7a (red) and DAPI (blue, nuclei). Brg1 was sill detected in P1 Atoh1-Brg1 −/− HCs with a lower signal than in control HCs while in P4 Atoh1-Brg1 −/− HCs, Brg1 was absent in all HCs. Scale bars: 20 μm.
    Rabbit Anti Brg1, supplied by Abcam, used in various techniques. Bioz Stars score: 92/100, based on 29 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    78
    Abcam anti smarcc1 baf155
    <t>Brg1</t> expression in cochlea and conditional Brg1 inactivation in cochlear HCs. ( A ) Transverse sections of P4 cochlea stained with Brg1 (green), the OHC marker Prestin (red) and DAPI (blue, nuclei). Brg1 was specifically deleted in Atoh1-Brg1 −/− HCs. Inserts represent higher magnification views of the boxed areas. Brackets indicate OHCs, and arrowheads indicate <t>IHC.</t> Abbreviations: RM, Reissner’s membrane; SV, stria vascularis; OC, organ of Corti; SG, spiral ganglia. Scale bars: 100 μm. ( B ) Efficiency of Brg1 deletion shown by whole mount cochlea of P1 and P4 mice stained with Brg1 (green), the HC marker Myosin7a (red) and DAPI (blue, nuclei). Brg1 was sill detected in P1 Atoh1-Brg1 −/− HCs with a lower signal than in control HCs while in P4 Atoh1-Brg1 −/− HCs, Brg1 was absent in all HCs. Scale bars: 20 μm.
    Anti Smarcc1 Baf155, supplied by Abcam, used in various techniques. Bioz Stars score: 78/100, based on 4 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    90
    Abcam rabbit anti baf155
    <t>Brg1</t> expression in cochlea and conditional Brg1 inactivation in cochlear HCs. ( A ) Transverse sections of P4 cochlea stained with Brg1 (green), the OHC marker Prestin (red) and DAPI (blue, nuclei). Brg1 was specifically deleted in Atoh1-Brg1 −/− HCs. Inserts represent higher magnification views of the boxed areas. Brackets indicate OHCs, and arrowheads indicate <t>IHC.</t> Abbreviations: RM, Reissner’s membrane; SV, stria vascularis; OC, organ of Corti; SG, spiral ganglia. Scale bars: 100 μm. ( B ) Efficiency of Brg1 deletion shown by whole mount cochlea of P1 and P4 mice stained with Brg1 (green), the HC marker Myosin7a (red) and DAPI (blue, nuclei). Brg1 was sill detected in P1 Atoh1-Brg1 −/− HCs with a lower signal than in control HCs while in P4 Atoh1-Brg1 −/− HCs, Brg1 was absent in all HCs. Scale bars: 20 μm.
    Rabbit Anti Baf155, supplied by Abcam, used in various techniques. Bioz Stars score: 90/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    81
    Abcam anti baf155
    <t>Brg1</t> expression in cochlea and conditional Brg1 inactivation in cochlear HCs. ( A ) Transverse sections of P4 cochlea stained with Brg1 (green), the OHC marker Prestin (red) and DAPI (blue, nuclei). Brg1 was specifically deleted in Atoh1-Brg1 −/− HCs. Inserts represent higher magnification views of the boxed areas. Brackets indicate OHCs, and arrowheads indicate <t>IHC.</t> Abbreviations: RM, Reissner’s membrane; SV, stria vascularis; OC, organ of Corti; SG, spiral ganglia. Scale bars: 100 μm. ( B ) Efficiency of Brg1 deletion shown by whole mount cochlea of P1 and P4 mice stained with Brg1 (green), the HC marker Myosin7a (red) and DAPI (blue, nuclei). Brg1 was sill detected in P1 Atoh1-Brg1 −/− HCs with a lower signal than in control HCs while in P4 Atoh1-Brg1 −/− HCs, Brg1 was absent in all HCs. Scale bars: 20 μm.
    Anti Baf155, supplied by Abcam, used in various techniques. Bioz Stars score: 81/100, based on 10 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Image Search Results


    The PTEN/AKT/GSK3β axis modulates BRG1 stability through the FBXW7-dependent ubiquitin proteasome pathway. ( A ) Sequence alignment of the putative GSK3β phosphorylation sites at S1417 and S1421 of BRG1. ( B ) In vitro kinase assays depicting major GSK3β phosphorylation sites in BRG1. ( C ) IB analysis of WCL and immunoprecipitates from control and PTEN-overexpressing PC3 cells treated with CHIR-99021 or λ-phosphatase as indicated. ( D ) Lysates from control and FBXW7-overexpressing cells were subjected to IP with an anti-BRG1 antibody, and ubiquitinated BRG1 was detected by an anti-Ub antibody. ( E ) IB analysis of PC3 cells transfected with scramble or AKT oligonucleotides with or without FBXW7 KD (shFBXW7). ( F ) Flag-tagged WT, BRG1-SA, and BRG1-SD proteins were incubated with SCF-FBXW7 complex as indicated and then subjected to Western blotting. ( G ) IB analysis of the indicated protein in WCL and immunoprecipitates from 293T cells transfected with HA-tagged FBXW7 and Flag-tagged BRG1, BRG-SA, or BRG1-SD. ( H ) IB analysis of the indicated protein in WT, SA, and SD cells. ( I ) WT, SA, and SD cell lysates were subjected to IP with the indicated antibodies. ( J ) Volume of subcutaneous tumors derived from WT, SA, and SD cells ( n = 6, 2-way ANOVA followed by Tukey’s multiple comparisons test). Scale bar: 1 cm. ( K ) Representative image of BRG1, P-1417/1421, and PTEN expression in lysates from PCa samples (upper panel). Pearson’s correlations among BRG1, P-1417/1421, and PTEN in PCa specimens are summarized in the heatmap ( n = 30). ** P

    Journal: The Journal of Clinical Investigation

    Article Title: Chromatin remodeling ATPase BRG1 and PTEN are synthetic lethal in prostate cancer

    doi: 10.1172/JCI123557

    Figure Lengend Snippet: The PTEN/AKT/GSK3β axis modulates BRG1 stability through the FBXW7-dependent ubiquitin proteasome pathway. ( A ) Sequence alignment of the putative GSK3β phosphorylation sites at S1417 and S1421 of BRG1. ( B ) In vitro kinase assays depicting major GSK3β phosphorylation sites in BRG1. ( C ) IB analysis of WCL and immunoprecipitates from control and PTEN-overexpressing PC3 cells treated with CHIR-99021 or λ-phosphatase as indicated. ( D ) Lysates from control and FBXW7-overexpressing cells were subjected to IP with an anti-BRG1 antibody, and ubiquitinated BRG1 was detected by an anti-Ub antibody. ( E ) IB analysis of PC3 cells transfected with scramble or AKT oligonucleotides with or without FBXW7 KD (shFBXW7). ( F ) Flag-tagged WT, BRG1-SA, and BRG1-SD proteins were incubated with SCF-FBXW7 complex as indicated and then subjected to Western blotting. ( G ) IB analysis of the indicated protein in WCL and immunoprecipitates from 293T cells transfected with HA-tagged FBXW7 and Flag-tagged BRG1, BRG-SA, or BRG1-SD. ( H ) IB analysis of the indicated protein in WT, SA, and SD cells. ( I ) WT, SA, and SD cell lysates were subjected to IP with the indicated antibodies. ( J ) Volume of subcutaneous tumors derived from WT, SA, and SD cells ( n = 6, 2-way ANOVA followed by Tukey’s multiple comparisons test). Scale bar: 1 cm. ( K ) Representative image of BRG1, P-1417/1421, and PTEN expression in lysates from PCa samples (upper panel). Pearson’s correlations among BRG1, P-1417/1421, and PTEN in PCa specimens are summarized in the heatmap ( n = 30). ** P

    Article Snippet: For the ChIP-Seq assay, chromatin was prepared from 3 biological replicates of control and PTEN-deleted 22RV-1 cells, and ChIP-Seq assays were then performed by Active Motif Inc. using an antibody against BRG1 (Abcam, catalog ab110641).

    Techniques: Sequencing, In Vitro, Transfection, Incubation, Western Blot, Derivative Assay, Expressing

    Targeting the SWI/SNF remodeling complex inhibits the progression of PTEN-deficient PCa. ( A ) Relative growth of PTEN-WT and PTEN-KD 22RV-1 cells treated with PFI-3 at different concentrations, as indicated. ( B ) Effects of PFI-3 treatment (50 mg/kg, once per week) on xenografts, as indicated ( n = 6 per group, 2-way ANOVA followed by Tukey’s multiple comparisons test). Treatment started when tumors reached 50–100 mm 3 . ( C and D ) Representative images of PTEN-deficient ( C ) or Myc-overexpressing ( D ) organoids treated with vehicle or PFI-3 (100 nM; 7 days). Scale bar: 200 μm. ( E ) MRI analysis of prostates in Pten PC–/– mice treated with PFI-3 for 45 days (50 mg/kg, starting at 2.5 months of age; T0). Prostate tumors are indicated by red dotted circles, and relative tumor volume is shown at the bottom ( n = 5, 2-tailed Student’s t test). Red asterisks indicate bladders. ( F ) H E staining of prostates from vehicle- and PFI-3-treated Pten PC–/– mice. Histology quantitation is indicated at the bottom ( n = 5, χ 2 test). Scale bar: 100 μm. ( G ) PTEN loss stabilized BRG1 through the inhibition of the AKT/GSK3β/FBXW7-mediated proteasome pathway. Consequently, BRG1 remodeled the chromatin configuration and initiated a PTEN-dependent BRG1 transcriptome to sustain tumor cell growth. Thus, targeting BRG1 represents a promising approach against PTEN-mutated prostate tumors. n = 3 independent experiments, 2-tailed Student’s t test ( A , C , and D ). * P

    Journal: The Journal of Clinical Investigation

    Article Title: Chromatin remodeling ATPase BRG1 and PTEN are synthetic lethal in prostate cancer

    doi: 10.1172/JCI123557

    Figure Lengend Snippet: Targeting the SWI/SNF remodeling complex inhibits the progression of PTEN-deficient PCa. ( A ) Relative growth of PTEN-WT and PTEN-KD 22RV-1 cells treated with PFI-3 at different concentrations, as indicated. ( B ) Effects of PFI-3 treatment (50 mg/kg, once per week) on xenografts, as indicated ( n = 6 per group, 2-way ANOVA followed by Tukey’s multiple comparisons test). Treatment started when tumors reached 50–100 mm 3 . ( C and D ) Representative images of PTEN-deficient ( C ) or Myc-overexpressing ( D ) organoids treated with vehicle or PFI-3 (100 nM; 7 days). Scale bar: 200 μm. ( E ) MRI analysis of prostates in Pten PC–/– mice treated with PFI-3 for 45 days (50 mg/kg, starting at 2.5 months of age; T0). Prostate tumors are indicated by red dotted circles, and relative tumor volume is shown at the bottom ( n = 5, 2-tailed Student’s t test). Red asterisks indicate bladders. ( F ) H E staining of prostates from vehicle- and PFI-3-treated Pten PC–/– mice. Histology quantitation is indicated at the bottom ( n = 5, χ 2 test). Scale bar: 100 μm. ( G ) PTEN loss stabilized BRG1 through the inhibition of the AKT/GSK3β/FBXW7-mediated proteasome pathway. Consequently, BRG1 remodeled the chromatin configuration and initiated a PTEN-dependent BRG1 transcriptome to sustain tumor cell growth. Thus, targeting BRG1 represents a promising approach against PTEN-mutated prostate tumors. n = 3 independent experiments, 2-tailed Student’s t test ( A , C , and D ). * P

    Article Snippet: For the ChIP-Seq assay, chromatin was prepared from 3 biological replicates of control and PTEN-deleted 22RV-1 cells, and ChIP-Seq assays were then performed by Active Motif Inc. using an antibody against BRG1 (Abcam, catalog ab110641).

    Techniques: Magnetic Resonance Imaging, Mouse Assay, Staining, Quantitation Assay, Inhibition

    BRG1 modulates c-Myc and MAPK signaling in PTEN-deficient cells. ( A ) RT-qPCR analysis of gene expressions in PTEN-KD and PTEN; BRG1-KD cells. ( B ) ChIP-qPCR assay of BRG1 binding, H3K27ac, and H3K27me3 in genes, as indicated. Arrows indicate primer locations. ( C ) IB analysis of the indicated protein in PTEN-KD and PTEN; BRG1-KD cells. ( D ) IB analysis of p-ERK levels in response to FGF treatment in control and PTEN-KD 22RV-1 cells with or without BRG1 KD (shBRG1). ( E ) IHC staining of c-Myc and p-ERK in prostate sections from Pten PC–/– , and Pten PC–/– ; Brg1 PC–/– mice. Scale bar: 50 μm. Quantitative data from 3 independent experiments, 2-tailed Student’s t test ( A and B ). * P

    Journal: The Journal of Clinical Investigation

    Article Title: Chromatin remodeling ATPase BRG1 and PTEN are synthetic lethal in prostate cancer

    doi: 10.1172/JCI123557

    Figure Lengend Snippet: BRG1 modulates c-Myc and MAPK signaling in PTEN-deficient cells. ( A ) RT-qPCR analysis of gene expressions in PTEN-KD and PTEN; BRG1-KD cells. ( B ) ChIP-qPCR assay of BRG1 binding, H3K27ac, and H3K27me3 in genes, as indicated. Arrows indicate primer locations. ( C ) IB analysis of the indicated protein in PTEN-KD and PTEN; BRG1-KD cells. ( D ) IB analysis of p-ERK levels in response to FGF treatment in control and PTEN-KD 22RV-1 cells with or without BRG1 KD (shBRG1). ( E ) IHC staining of c-Myc and p-ERK in prostate sections from Pten PC–/– , and Pten PC–/– ; Brg1 PC–/– mice. Scale bar: 50 μm. Quantitative data from 3 independent experiments, 2-tailed Student’s t test ( A and B ). * P

    Article Snippet: For the ChIP-Seq assay, chromatin was prepared from 3 biological replicates of control and PTEN-deleted 22RV-1 cells, and ChIP-Seq assays were then performed by Active Motif Inc. using an antibody against BRG1 (Abcam, catalog ab110641).

    Techniques: Quantitative RT-PCR, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Binding Assay, Immunohistochemistry, Staining, Mouse Assay

    BRG1 is required in PTEN-deficient PCa cells. ( A ) MTT analysis of PCa cells with or without BRG1 KD (shBRG1). ( B ) Transwell (upper right) and soft agar (lower right) images of BRG1-KD PC3 cells with or without WT or mutant BRG1 (K798R) restoration. Scale bar: 1 mm. ( C ) IB of lysates and cell growth measurements in control and BRG1-KD 22RV-1 and LAPC4 cells with or without PTEN KD (shPTEN). ( D ) Measurement of subcutaneous tumor growth of control and PTEN-KD 22RV-1 cells with or without BRG1 depletion (shBRG1) ( n = 6, 2-way ANOVA followed by Tukey’s multiple comparisons test); a representative image is shown. Scale bar: 1 cm. ( E ) Representative BLI images for control and BRG1-KD PC3 cells at day 0 (upper panels) and day 60 (lower panels). Limb metastasis is calculated as the mean ± SEM of the bioluminescence signal at day 60 ( n = 6 per group, 2-tailed Student’s t test). ( F ) Representative x-ray images of bone metastasis are shown on the left, and the osteolytic area is quantified on the right ( n = 8, 2-tailed Student’s t test). ( G ) TRAP- and E-cadherin–stained images as indicated. T, tumor cell; M, bone marrow; arrow, TRAP-positive cell. Scale bar: 50 μm. Data represent mean ± SEM of 3 independent experiments. Statistical analyses were performed by 2-way ANOVA followed by Tukey’s multiple comparisons test ( A and C ). ** P

    Journal: The Journal of Clinical Investigation

    Article Title: Chromatin remodeling ATPase BRG1 and PTEN are synthetic lethal in prostate cancer

    doi: 10.1172/JCI123557

    Figure Lengend Snippet: BRG1 is required in PTEN-deficient PCa cells. ( A ) MTT analysis of PCa cells with or without BRG1 KD (shBRG1). ( B ) Transwell (upper right) and soft agar (lower right) images of BRG1-KD PC3 cells with or without WT or mutant BRG1 (K798R) restoration. Scale bar: 1 mm. ( C ) IB of lysates and cell growth measurements in control and BRG1-KD 22RV-1 and LAPC4 cells with or without PTEN KD (shPTEN). ( D ) Measurement of subcutaneous tumor growth of control and PTEN-KD 22RV-1 cells with or without BRG1 depletion (shBRG1) ( n = 6, 2-way ANOVA followed by Tukey’s multiple comparisons test); a representative image is shown. Scale bar: 1 cm. ( E ) Representative BLI images for control and BRG1-KD PC3 cells at day 0 (upper panels) and day 60 (lower panels). Limb metastasis is calculated as the mean ± SEM of the bioluminescence signal at day 60 ( n = 6 per group, 2-tailed Student’s t test). ( F ) Representative x-ray images of bone metastasis are shown on the left, and the osteolytic area is quantified on the right ( n = 8, 2-tailed Student’s t test). ( G ) TRAP- and E-cadherin–stained images as indicated. T, tumor cell; M, bone marrow; arrow, TRAP-positive cell. Scale bar: 50 μm. Data represent mean ± SEM of 3 independent experiments. Statistical analyses were performed by 2-way ANOVA followed by Tukey’s multiple comparisons test ( A and C ). ** P

    Article Snippet: For the ChIP-Seq assay, chromatin was prepared from 3 biological replicates of control and PTEN-deleted 22RV-1 cells, and ChIP-Seq assays were then performed by Active Motif Inc. using an antibody against BRG1 (Abcam, catalog ab110641).

    Techniques: MTT Assay, Mutagenesis, Staining

    Brg1 loss inhibits Pten loss–induced tumorigenesis in mice. ( A ) IHC staining of BRG1 in prostate sections of 5-month-old Pten PC–/– , and Pten PC–/– ; Brg1 PC–/– mice. Scale bar: 50 μm. ( B ) Representative BLI images of Pten PC–/– , and Pten PC–/– ; Brg1 PC–/– mice at 5 months of age. Quantification of BLI is shown on the right ( n = 5, 2-tailed Student’s t test). ( C ) H E-stained sections of representative anterior prostate (AP), dorsal-lateral prostate (DLP), and ventral prostate (VP). Scale bars: 100 μm. Original magnification for the inset, × 50. ( D ) Quantification of histology grade in Pten PC–/– ( n = 10), and Pten PC–/– ; Brg1 PC–/– mice ( n = 6) at 5 months of age (χ 2 test). ( E ) H E, AR, and SMAα staining of the DLP in Pten PC–/– (with the appearance of invasive adenocarcinoma) and Pten PC–/– ; Brg1 PC–/– mice. Scale bars: 100 μm. ( F ) Representative images of organoids from Pten-WT and Pten-null prostates with or without BRG1 KD (shBRG1). Quantitation of organoid size is representative of 3 experiments shown on the right (2-tailed Student’s t test). Scale bar: 200 μm. ( G ) Immunostaining of Ki67, AR, and P63 as indicated. Scale bar: 50 μm. ( H ) Organoid images derived from prostatic Myc-overexpressing mice (Hi-Myc) with or without BRG1 KD (shBRG1); quantitative results are representative of 3 experiments shown at the bottom (2-tailed Student’s t test). Scale bar: 400 μm. ** P

    Journal: The Journal of Clinical Investigation

    Article Title: Chromatin remodeling ATPase BRG1 and PTEN are synthetic lethal in prostate cancer

    doi: 10.1172/JCI123557

    Figure Lengend Snippet: Brg1 loss inhibits Pten loss–induced tumorigenesis in mice. ( A ) IHC staining of BRG1 in prostate sections of 5-month-old Pten PC–/– , and Pten PC–/– ; Brg1 PC–/– mice. Scale bar: 50 μm. ( B ) Representative BLI images of Pten PC–/– , and Pten PC–/– ; Brg1 PC–/– mice at 5 months of age. Quantification of BLI is shown on the right ( n = 5, 2-tailed Student’s t test). ( C ) H E-stained sections of representative anterior prostate (AP), dorsal-lateral prostate (DLP), and ventral prostate (VP). Scale bars: 100 μm. Original magnification for the inset, × 50. ( D ) Quantification of histology grade in Pten PC–/– ( n = 10), and Pten PC–/– ; Brg1 PC–/– mice ( n = 6) at 5 months of age (χ 2 test). ( E ) H E, AR, and SMAα staining of the DLP in Pten PC–/– (with the appearance of invasive adenocarcinoma) and Pten PC–/– ; Brg1 PC–/– mice. Scale bars: 100 μm. ( F ) Representative images of organoids from Pten-WT and Pten-null prostates with or without BRG1 KD (shBRG1). Quantitation of organoid size is representative of 3 experiments shown on the right (2-tailed Student’s t test). Scale bar: 200 μm. ( G ) Immunostaining of Ki67, AR, and P63 as indicated. Scale bar: 50 μm. ( H ) Organoid images derived from prostatic Myc-overexpressing mice (Hi-Myc) with or without BRG1 KD (shBRG1); quantitative results are representative of 3 experiments shown at the bottom (2-tailed Student’s t test). Scale bar: 400 μm. ** P

    Article Snippet: For the ChIP-Seq assay, chromatin was prepared from 3 biological replicates of control and PTEN-deleted 22RV-1 cells, and ChIP-Seq assays were then performed by Active Motif Inc. using an antibody against BRG1 (Abcam, catalog ab110641).

    Techniques: Mouse Assay, Immunohistochemistry, Staining, Quantitation Assay, Immunostaining, Derivative Assay

    Identification of the epigenetic regulator required for PTEN-deficient PCa cells. ( A ) A schematic of the screening workflow for the chromatin regulator based on CRISPR-Cas9 screening in PTEN-WT and PTEN-KD (shPTEN) 22RV-1 cells. ( B ) Scatter plot showing the normalized counts for each sgRNA in the original pool (day 0) relative to the samples taken after 45 days of cultures (MaGeCK and MAGeCK-VISPR analysis). ( C ) Venn diagram showing the number of overlapping genes between the cells as indicated. ( D ) siRNA KD of 32 candidate genes and their effects on the growth of PTEN-WT and PTEN-KD 22RV-1 cells. Quantitative results shown are representative of 4 experiments. Genes highlighted in red box exhibited the differential growth effects between PTEN-WT and PTEN-KD 22RV-1 cells. ( E ) BRG1 staining indexes using a 10-point quantification scale in cohorts of normal prostate tissue ( n = 87) and prostate tumors ( n = 122) (Wilcoxon’s rank sum test). Scale bar: 50 μm. ( F ) Kaplan-Meier plot of recurrence after radical prostatectomy based on the BRG1 expression index in patients ( P values by log-rank test). Scale bar: 200 μm. ( G ) Kaplan-Meier plots based on BRG1 expression in PTEN-low and PTEN-high tumors (log-rank test).

    Journal: The Journal of Clinical Investigation

    Article Title: Chromatin remodeling ATPase BRG1 and PTEN are synthetic lethal in prostate cancer

    doi: 10.1172/JCI123557

    Figure Lengend Snippet: Identification of the epigenetic regulator required for PTEN-deficient PCa cells. ( A ) A schematic of the screening workflow for the chromatin regulator based on CRISPR-Cas9 screening in PTEN-WT and PTEN-KD (shPTEN) 22RV-1 cells. ( B ) Scatter plot showing the normalized counts for each sgRNA in the original pool (day 0) relative to the samples taken after 45 days of cultures (MaGeCK and MAGeCK-VISPR analysis). ( C ) Venn diagram showing the number of overlapping genes between the cells as indicated. ( D ) siRNA KD of 32 candidate genes and their effects on the growth of PTEN-WT and PTEN-KD 22RV-1 cells. Quantitative results shown are representative of 4 experiments. Genes highlighted in red box exhibited the differential growth effects between PTEN-WT and PTEN-KD 22RV-1 cells. ( E ) BRG1 staining indexes using a 10-point quantification scale in cohorts of normal prostate tissue ( n = 87) and prostate tumors ( n = 122) (Wilcoxon’s rank sum test). Scale bar: 50 μm. ( F ) Kaplan-Meier plot of recurrence after radical prostatectomy based on the BRG1 expression index in patients ( P values by log-rank test). Scale bar: 200 μm. ( G ) Kaplan-Meier plots based on BRG1 expression in PTEN-low and PTEN-high tumors (log-rank test).

    Article Snippet: For the ChIP-Seq assay, chromatin was prepared from 3 biological replicates of control and PTEN-deleted 22RV-1 cells, and ChIP-Seq assays were then performed by Active Motif Inc. using an antibody against BRG1 (Abcam, catalog ab110641).

    Techniques: CRISPR, Staining, Expressing

    Brg1 expression in cochlea and conditional Brg1 inactivation in cochlear HCs. ( A ) Transverse sections of P4 cochlea stained with Brg1 (green), the OHC marker Prestin (red) and DAPI (blue, nuclei). Brg1 was specifically deleted in Atoh1-Brg1 −/− HCs. Inserts represent higher magnification views of the boxed areas. Brackets indicate OHCs, and arrowheads indicate IHC. Abbreviations: RM, Reissner’s membrane; SV, stria vascularis; OC, organ of Corti; SG, spiral ganglia. Scale bars: 100 μm. ( B ) Efficiency of Brg1 deletion shown by whole mount cochlea of P1 and P4 mice stained with Brg1 (green), the HC marker Myosin7a (red) and DAPI (blue, nuclei). Brg1 was sill detected in P1 Atoh1-Brg1 −/− HCs with a lower signal than in control HCs while in P4 Atoh1-Brg1 −/− HCs, Brg1 was absent in all HCs. Scale bars: 20 μm.

    Journal: Scientific Reports

    Article Title: Deletion of Brg1 causes abnormal hair cell planer polarity, hair cell anchorage, and scar formation in mouse cochlea

    doi: 10.1038/srep27124

    Figure Lengend Snippet: Brg1 expression in cochlea and conditional Brg1 inactivation in cochlear HCs. ( A ) Transverse sections of P4 cochlea stained with Brg1 (green), the OHC marker Prestin (red) and DAPI (blue, nuclei). Brg1 was specifically deleted in Atoh1-Brg1 −/− HCs. Inserts represent higher magnification views of the boxed areas. Brackets indicate OHCs, and arrowheads indicate IHC. Abbreviations: RM, Reissner’s membrane; SV, stria vascularis; OC, organ of Corti; SG, spiral ganglia. Scale bars: 100 μm. ( B ) Efficiency of Brg1 deletion shown by whole mount cochlea of P1 and P4 mice stained with Brg1 (green), the HC marker Myosin7a (red) and DAPI (blue, nuclei). Brg1 was sill detected in P1 Atoh1-Brg1 −/− HCs with a lower signal than in control HCs while in P4 Atoh1-Brg1 −/− HCs, Brg1 was absent in all HCs. Scale bars: 20 μm.

    Article Snippet: The following primary antibodies were used for immunostaining: anti-Brg1 (rabbit, 1:400, Abcam), anti-Myosin7a (rabbit, 1:400, Proteus Biosciences), anti-Myosin7a (goat, 1:800, Santa Cruz), anti-LGN (rabbit, 1:200, Proteintech), anti-Gαi3 (rabbit, 1:400, Sigma), anti-acetylated-α-tubulin (mouse, 1:400, Sigma), anti-α-tubulin (rabbit, 1:200, Proteintech), anti-βII-Spectrin (mouse, 1:400, BD), anti-Fz6 (goat, 1:500, RD), anti-Vangl1 (rabbit, 1:500, Sigma), anti-Prestin (goat, 1:400, Santa Cruz), anti-aPKC (rabbit, 1:200, Santa Cruz), anti-Cleaved-Caspase3 (rabbit, 1:400, CST), anti-ZO-1 (rabbit, 1:400, Invitrogen), anti-E-Cadherin (rabbit, 1:200, CST), anti-β-Catenin (rabbit, 1:400, Abcam), anti-Cyclin D1 (rabbit, 1:400, Abcam), anti-Ki67 (rabbit, 1:400, Abcam), anti-PCNA (rabbit, 1:400, Abcam), anti-Phospho-Histone3 (PH3) (rabbit, 1:800, Bioworld), anti-BrdU (mouse, 1:1000, Sigma), anti-Par3 (rabbit, 1:200, Proteintech).

    Techniques: Expressing, Staining, Marker, Immunohistochemistry, Mouse Assay

    Broken reticular lamina after HC loss in Atoh1-Brg1 −/− mice and HC loss rescue in vitro . ( A ) Whole-mount cochlea stained with ZO-1 (green) and the HC marker Myosin7a (red). ZO-1 shown the same expression pattern in the control and Atoh1-Brg1 −/− auditory epithelia at P8. In P14 auditory epithelia, large areas without ZO-1 appeared at some sites where HC loss occurred. Asterisks indicate large ZO-1 free areas. Scale bar: 20 μm. ( B ) SEM images showing the holes in the P14 Atoh1-Brg1 −/− reticular lamina formed by the collapsed HC cuticular plate. Arrows indicate holes in the reticular lamina. Scale bars: 5 μm. ( C ) Whole-mount cochlea of explants of Atoh1-Brg1 −/− and control mice and of Atoh1-Brg1 −/− and control mice in vivo stained with the HC marker Myosin7a. Upper panel shows the cochlea specimen from P20 mice. The lower panel shows cochlea explants from the middle turn of the cochlea maintained in normal culture conditions from P5 to P20. Almost all of the IHCs survive in Atoh1-Brg1 −/− cochlea explants, and more OHCs survive compared with the Atoh1-Brg1 −/− cochlea from P20 mice in vivo . Scale bar: 50 μm. ( D ) Quantification of IHC number in the cochlea of explants of Atoh1-Brg1 −/− and control mice and of Atoh1-Brg1 −/− and control mice (p = 0.0089). The error bars indicate the SEM. **P

    Journal: Scientific Reports

    Article Title: Deletion of Brg1 causes abnormal hair cell planer polarity, hair cell anchorage, and scar formation in mouse cochlea

    doi: 10.1038/srep27124

    Figure Lengend Snippet: Broken reticular lamina after HC loss in Atoh1-Brg1 −/− mice and HC loss rescue in vitro . ( A ) Whole-mount cochlea stained with ZO-1 (green) and the HC marker Myosin7a (red). ZO-1 shown the same expression pattern in the control and Atoh1-Brg1 −/− auditory epithelia at P8. In P14 auditory epithelia, large areas without ZO-1 appeared at some sites where HC loss occurred. Asterisks indicate large ZO-1 free areas. Scale bar: 20 μm. ( B ) SEM images showing the holes in the P14 Atoh1-Brg1 −/− reticular lamina formed by the collapsed HC cuticular plate. Arrows indicate holes in the reticular lamina. Scale bars: 5 μm. ( C ) Whole-mount cochlea of explants of Atoh1-Brg1 −/− and control mice and of Atoh1-Brg1 −/− and control mice in vivo stained with the HC marker Myosin7a. Upper panel shows the cochlea specimen from P20 mice. The lower panel shows cochlea explants from the middle turn of the cochlea maintained in normal culture conditions from P5 to P20. Almost all of the IHCs survive in Atoh1-Brg1 −/− cochlea explants, and more OHCs survive compared with the Atoh1-Brg1 −/− cochlea from P20 mice in vivo . Scale bar: 50 μm. ( D ) Quantification of IHC number in the cochlea of explants of Atoh1-Brg1 −/− and control mice and of Atoh1-Brg1 −/− and control mice (p = 0.0089). The error bars indicate the SEM. **P

    Article Snippet: The following primary antibodies were used for immunostaining: anti-Brg1 (rabbit, 1:400, Abcam), anti-Myosin7a (rabbit, 1:400, Proteus Biosciences), anti-Myosin7a (goat, 1:800, Santa Cruz), anti-LGN (rabbit, 1:200, Proteintech), anti-Gαi3 (rabbit, 1:400, Sigma), anti-acetylated-α-tubulin (mouse, 1:400, Sigma), anti-α-tubulin (rabbit, 1:200, Proteintech), anti-βII-Spectrin (mouse, 1:400, BD), anti-Fz6 (goat, 1:500, RD), anti-Vangl1 (rabbit, 1:500, Sigma), anti-Prestin (goat, 1:400, Santa Cruz), anti-aPKC (rabbit, 1:200, Santa Cruz), anti-Cleaved-Caspase3 (rabbit, 1:400, CST), anti-ZO-1 (rabbit, 1:400, Invitrogen), anti-E-Cadherin (rabbit, 1:200, CST), anti-β-Catenin (rabbit, 1:400, Abcam), anti-Cyclin D1 (rabbit, 1:400, Abcam), anti-Ki67 (rabbit, 1:400, Abcam), anti-PCNA (rabbit, 1:400, Abcam), anti-Phospho-Histone3 (PH3) (rabbit, 1:800, Bioworld), anti-BrdU (mouse, 1:1000, Sigma), anti-Par3 (rabbit, 1:200, Proteintech).

    Techniques: Mouse Assay, In Vitro, Staining, Marker, Expressing, In Vivo, Immunohistochemistry