brg1 antibody  (Abcam)

 
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    Name:
    Anti BRG1 antibody
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    Catalog Number:
    ab4081
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    Structured Review

    Abcam brg1 antibody
    <t>BRG1</t> plays a tumor suppressor role. (A) Colony formation of 5637-NC and 5637-iBRG1 cells. The cells were cultured for 14 days. Colonies were stained using crystal violet. (B) Colony formation of 5637-pcDNA3.1 and 5637-BRG1 cells. The cells were cultured for 14 days. Colonies were stained using crystal violet. (C) Growth curves of 5637 cells after transfection with BRG1 RNAi. Cellular proliferation was measured using MTT assay at 24, 48, 72, 96 and 120 h. The differences in data of 72, 96 and 120 h were significant, * P

    https://www.bioz.com/result/brg1 antibody/product/Abcam
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    brg1 antibody - by Bioz Stars, 2020-09
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    Images

    1) Product Images from "Long non-coding RNA urothelial carcinoma associated 1 induces cell replication by inhibiting BRG1 in 5637 cells"

    Article Title: Long non-coding RNA urothelial carcinoma associated 1 induces cell replication by inhibiting BRG1 in 5637 cells

    Journal: Oncology Reports

    doi: 10.3892/or.2014.3309

    BRG1 plays a tumor suppressor role. (A) Colony formation of 5637-NC and 5637-iBRG1 cells. The cells were cultured for 14 days. Colonies were stained using crystal violet. (B) Colony formation of 5637-pcDNA3.1 and 5637-BRG1 cells. The cells were cultured for 14 days. Colonies were stained using crystal violet. (C) Growth curves of 5637 cells after transfection with BRG1 RNAi. Cellular proliferation was measured using MTT assay at 24, 48, 72, 96 and 120 h. The differences in data of 72, 96 and 120 h were significant, * P
    Figure Legend Snippet: BRG1 plays a tumor suppressor role. (A) Colony formation of 5637-NC and 5637-iBRG1 cells. The cells were cultured for 14 days. Colonies were stained using crystal violet. (B) Colony formation of 5637-pcDNA3.1 and 5637-BRG1 cells. The cells were cultured for 14 days. Colonies were stained using crystal violet. (C) Growth curves of 5637 cells after transfection with BRG1 RNAi. Cellular proliferation was measured using MTT assay at 24, 48, 72, 96 and 120 h. The differences in data of 72, 96 and 120 h were significant, * P

    Techniques Used: Cell Culture, Staining, Transfection, MTT Assay

    UCA1 blocks recruitment of BRG1 to chromatin. (A) UCA1 does not affect the ATPase activity of BRG1. The kinetics of BRG1-induced ATP hydrolysis were analyzed in the presence or absence of UCA1. (B) ChIP analysis of BRG1 binding to the p21 promoter in 5637-iUCA1. 5637-NC cells were used as the control. Genomic DNA was fixed and immunoprecipitated using anti-BRG1 antibody, with IgG as a negative control. Real-time PCR was performed using a primer set specific to the BRG1-binding site of p21 promoter. Data were normalized to input and are expressed as the means ± SD of three independent experiments. * P
    Figure Legend Snippet: UCA1 blocks recruitment of BRG1 to chromatin. (A) UCA1 does not affect the ATPase activity of BRG1. The kinetics of BRG1-induced ATP hydrolysis were analyzed in the presence or absence of UCA1. (B) ChIP analysis of BRG1 binding to the p21 promoter in 5637-iUCA1. 5637-NC cells were used as the control. Genomic DNA was fixed and immunoprecipitated using anti-BRG1 antibody, with IgG as a negative control. Real-time PCR was performed using a primer set specific to the BRG1-binding site of p21 promoter. Data were normalized to input and are expressed as the means ± SD of three independent experiments. * P

    Techniques Used: Activity Assay, Chromatin Immunoprecipitation, Binding Assay, Immunoprecipitation, Negative Control, Real-time Polymerase Chain Reaction

    UCA1 binds to BRG1 in vitro and in vivo . (A) Electrophoresis of proteins bound to biotin-labeled sense and antisense UCA1 in NuPAGE 4–12% Bis-Tris gel. The biotin-labeled sense and antisense UCA1 were transcribed in vitro and incubated with HeLa cell lysate. The arrow shows the protein band for BRG1. (B) Western blot analysis of biotin-labeled sense and antisense UCA1-bound proteins using antibody against BRG1. (C) UCA1 binding BRG1 in 5637 cells was detected by RNA-binding protein immunoprecipitation assay. Antibody against BRG1 and mouse IgG (as a negative control) were used to pull-down RNAs in 5637 cells. The level of UCA1 was determined using real-time PCR. Data were normalized to input and are expressed as the means ± SD of three independent experiments * P
    Figure Legend Snippet: UCA1 binds to BRG1 in vitro and in vivo . (A) Electrophoresis of proteins bound to biotin-labeled sense and antisense UCA1 in NuPAGE 4–12% Bis-Tris gel. The biotin-labeled sense and antisense UCA1 were transcribed in vitro and incubated with HeLa cell lysate. The arrow shows the protein band for BRG1. (B) Western blot analysis of biotin-labeled sense and antisense UCA1-bound proteins using antibody against BRG1. (C) UCA1 binding BRG1 in 5637 cells was detected by RNA-binding protein immunoprecipitation assay. Antibody against BRG1 and mouse IgG (as a negative control) were used to pull-down RNAs in 5637 cells. The level of UCA1 was determined using real-time PCR. Data were normalized to input and are expressed as the means ± SD of three independent experiments * P

    Techniques Used: In Vitro, In Vivo, Electrophoresis, Labeling, Incubation, Western Blot, Binding Assay, RNA Binding Assay, Immunoprecipitation, Negative Control, Real-time Polymerase Chain Reaction

    UCA1 antagonizes the tumor suppressing function of BRG1. (A) (Left) Colony formation of EJ control, EJ-BRG1 (overexpressed BRG1) and EJ-BRG1-UCA1 (overexpressed BRG1 and UCA1) cells. The cells were cultured with G418 for 10 days. Colonies were stained using crystal violet. (Right) Colony formation of 5637 control, 5637-iUCA1 (UCA1 knocked down) and 5637-iUCA1-iBRG1 (UCA1 and BRG1 both knocked down) cells. The cells were cultured with G418 and puromycin for 10 days. Colonies were stained using crystal violet. (B) (Left) Growth curves of EJ control, EJ-BRG1 and EJ-BRG1-UCA1 cells. Cellular proliferation was measured using MTT assays at 24, 48, 72, 96 and 120 h. The differences between EJ-BRG1 and EJ-BRG1-UCA1 were significant. * P
    Figure Legend Snippet: UCA1 antagonizes the tumor suppressing function of BRG1. (A) (Left) Colony formation of EJ control, EJ-BRG1 (overexpressed BRG1) and EJ-BRG1-UCA1 (overexpressed BRG1 and UCA1) cells. The cells were cultured with G418 for 10 days. Colonies were stained using crystal violet. (Right) Colony formation of 5637 control, 5637-iUCA1 (UCA1 knocked down) and 5637-iUCA1-iBRG1 (UCA1 and BRG1 both knocked down) cells. The cells were cultured with G418 and puromycin for 10 days. Colonies were stained using crystal violet. (B) (Left) Growth curves of EJ control, EJ-BRG1 and EJ-BRG1-UCA1 cells. Cellular proliferation was measured using MTT assays at 24, 48, 72, 96 and 120 h. The differences between EJ-BRG1 and EJ-BRG1-UCA1 were significant. * P

    Techniques Used: Cell Culture, Staining, MTT Assay

    UCA1 expression correlates positively with BRG1 expression in bladder cancer tissue samples. (A) Real-time PCR was used to analyze UCA1 and BRG1 mRNA levels in the same samples. S, sample. (B) The status of UCA1 and BRG1 expression in bladder cancer specimens, > 2-fold was identified as high expression. The 2×2 correlation table and Fisher’s exact test were used. The two-sided value P=0.009 was considered significant. UCA1, urothelial carcinoma associated 1.
    Figure Legend Snippet: UCA1 expression correlates positively with BRG1 expression in bladder cancer tissue samples. (A) Real-time PCR was used to analyze UCA1 and BRG1 mRNA levels in the same samples. S, sample. (B) The status of UCA1 and BRG1 expression in bladder cancer specimens, > 2-fold was identified as high expression. The 2×2 correlation table and Fisher’s exact test were used. The two-sided value P=0.009 was considered significant. UCA1, urothelial carcinoma associated 1.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    2) Product Images from "The dilemma of early preventive oophorectomy in familial small cell carcinoma of the ovary of hypercalcemic type"

    Article Title: The dilemma of early preventive oophorectomy in familial small cell carcinoma of the ovary of hypercalcemic type

    Journal: Gynecologic Oncology Reports

    doi: 10.1016/j.gore.2019.02.002

    Retained SMARCA4 (BRG1) protein expression in one of the ovaries. There is positive nuclear staining of the granulosa cells of a developing follicle (right of photomicrograph) and the ovarian stroma (left of photomicrograph).
    Figure Legend Snippet: Retained SMARCA4 (BRG1) protein expression in one of the ovaries. There is positive nuclear staining of the granulosa cells of a developing follicle (right of photomicrograph) and the ovarian stroma (left of photomicrograph).

    Techniques Used: Expressing, Staining

    3) Product Images from "Long non-coding RNA urothelial carcinoma associated 1 induces cell replication by inhibiting BRG1 in 5637 cells"

    Article Title: Long non-coding RNA urothelial carcinoma associated 1 induces cell replication by inhibiting BRG1 in 5637 cells

    Journal: Oncology Reports

    doi: 10.3892/or.2014.3309

    BRG1 plays a tumor suppressor role. (A) Colony formation of 5637-NC and 5637-iBRG1 cells. The cells were cultured for 14 days. Colonies were stained using crystal violet. (B) Colony formation of 5637-pcDNA3.1 and 5637-BRG1 cells. The cells were cultured for 14 days. Colonies were stained using crystal violet. (C) Growth curves of 5637 cells after transfection with BRG1 RNAi. Cellular proliferation was measured using MTT assay at 24, 48, 72, 96 and 120 h. The differences in data of 72, 96 and 120 h were significant, * P
    Figure Legend Snippet: BRG1 plays a tumor suppressor role. (A) Colony formation of 5637-NC and 5637-iBRG1 cells. The cells were cultured for 14 days. Colonies were stained using crystal violet. (B) Colony formation of 5637-pcDNA3.1 and 5637-BRG1 cells. The cells were cultured for 14 days. Colonies were stained using crystal violet. (C) Growth curves of 5637 cells after transfection with BRG1 RNAi. Cellular proliferation was measured using MTT assay at 24, 48, 72, 96 and 120 h. The differences in data of 72, 96 and 120 h were significant, * P

    Techniques Used: Cell Culture, Staining, Transfection, MTT Assay

    UCA1 blocks recruitment of BRG1 to chromatin. (A) UCA1 does not affect the ATPase activity of BRG1. The kinetics of BRG1-induced ATP hydrolysis were analyzed in the presence or absence of UCA1. (B) ChIP analysis of BRG1 binding to the p21 promoter in 5637-iUCA1. 5637-NC cells were used as the control. Genomic DNA was fixed and immunoprecipitated using anti-BRG1 antibody, with IgG as a negative control. Real-time PCR was performed using a primer set specific to the BRG1-binding site of p21 promoter. Data were normalized to input and are expressed as the means ± SD of three independent experiments. * P
    Figure Legend Snippet: UCA1 blocks recruitment of BRG1 to chromatin. (A) UCA1 does not affect the ATPase activity of BRG1. The kinetics of BRG1-induced ATP hydrolysis were analyzed in the presence or absence of UCA1. (B) ChIP analysis of BRG1 binding to the p21 promoter in 5637-iUCA1. 5637-NC cells were used as the control. Genomic DNA was fixed and immunoprecipitated using anti-BRG1 antibody, with IgG as a negative control. Real-time PCR was performed using a primer set specific to the BRG1-binding site of p21 promoter. Data were normalized to input and are expressed as the means ± SD of three independent experiments. * P

    Techniques Used: Activity Assay, Chromatin Immunoprecipitation, Binding Assay, Immunoprecipitation, Negative Control, Real-time Polymerase Chain Reaction

    UCA1 binds to BRG1 in vitro and in vivo . (A) Electrophoresis of proteins bound to biotin-labeled sense and antisense UCA1 in NuPAGE 4–12% Bis-Tris gel. The biotin-labeled sense and antisense UCA1 were transcribed in vitro and incubated with HeLa cell lysate. The arrow shows the protein band for BRG1. (B) Western blot analysis of biotin-labeled sense and antisense UCA1-bound proteins using antibody against BRG1. (C) UCA1 binding BRG1 in 5637 cells was detected by RNA-binding protein immunoprecipitation assay. Antibody against BRG1 and mouse IgG (as a negative control) were used to pull-down RNAs in 5637 cells. The level of UCA1 was determined using real-time PCR. Data were normalized to input and are expressed as the means ± SD of three independent experiments * P
    Figure Legend Snippet: UCA1 binds to BRG1 in vitro and in vivo . (A) Electrophoresis of proteins bound to biotin-labeled sense and antisense UCA1 in NuPAGE 4–12% Bis-Tris gel. The biotin-labeled sense and antisense UCA1 were transcribed in vitro and incubated with HeLa cell lysate. The arrow shows the protein band for BRG1. (B) Western blot analysis of biotin-labeled sense and antisense UCA1-bound proteins using antibody against BRG1. (C) UCA1 binding BRG1 in 5637 cells was detected by RNA-binding protein immunoprecipitation assay. Antibody against BRG1 and mouse IgG (as a negative control) were used to pull-down RNAs in 5637 cells. The level of UCA1 was determined using real-time PCR. Data were normalized to input and are expressed as the means ± SD of three independent experiments * P

    Techniques Used: In Vitro, In Vivo, Electrophoresis, Labeling, Incubation, Western Blot, Binding Assay, RNA Binding Assay, Immunoprecipitation, Negative Control, Real-time Polymerase Chain Reaction

    UCA1 antagonizes the tumor suppressing function of BRG1. (A) (Left) Colony formation of EJ control, EJ-BRG1 (overexpressed BRG1) and EJ-BRG1-UCA1 (overexpressed BRG1 and UCA1) cells. The cells were cultured with G418 for 10 days. Colonies were stained using crystal violet. (Right) Colony formation of 5637 control, 5637-iUCA1 (UCA1 knocked down) and 5637-iUCA1-iBRG1 (UCA1 and BRG1 both knocked down) cells. The cells were cultured with G418 and puromycin for 10 days. Colonies were stained using crystal violet. (B) (Left) Growth curves of EJ control, EJ-BRG1 and EJ-BRG1-UCA1 cells. Cellular proliferation was measured using MTT assays at 24, 48, 72, 96 and 120 h. The differences between EJ-BRG1 and EJ-BRG1-UCA1 were significant. * P
    Figure Legend Snippet: UCA1 antagonizes the tumor suppressing function of BRG1. (A) (Left) Colony formation of EJ control, EJ-BRG1 (overexpressed BRG1) and EJ-BRG1-UCA1 (overexpressed BRG1 and UCA1) cells. The cells were cultured with G418 for 10 days. Colonies were stained using crystal violet. (Right) Colony formation of 5637 control, 5637-iUCA1 (UCA1 knocked down) and 5637-iUCA1-iBRG1 (UCA1 and BRG1 both knocked down) cells. The cells were cultured with G418 and puromycin for 10 days. Colonies were stained using crystal violet. (B) (Left) Growth curves of EJ control, EJ-BRG1 and EJ-BRG1-UCA1 cells. Cellular proliferation was measured using MTT assays at 24, 48, 72, 96 and 120 h. The differences between EJ-BRG1 and EJ-BRG1-UCA1 were significant. * P

    Techniques Used: Cell Culture, Staining, MTT Assay

    UCA1 expression correlates positively with BRG1 expression in bladder cancer tissue samples. (A) Real-time PCR was used to analyze UCA1 and BRG1 mRNA levels in the same samples. S, sample. (B) The status of UCA1 and BRG1 expression in bladder cancer specimens, > 2-fold was identified as high expression. The 2×2 correlation table and Fisher’s exact test were used. The two-sided value P=0.009 was considered significant. UCA1, urothelial carcinoma associated 1.
    Figure Legend Snippet: UCA1 expression correlates positively with BRG1 expression in bladder cancer tissue samples. (A) Real-time PCR was used to analyze UCA1 and BRG1 mRNA levels in the same samples. S, sample. (B) The status of UCA1 and BRG1 expression in bladder cancer specimens, > 2-fold was identified as high expression. The 2×2 correlation table and Fisher’s exact test were used. The two-sided value P=0.009 was considered significant. UCA1, urothelial carcinoma associated 1.

    Techniques Used: Expressing, Real-time Polymerase Chain Reaction

    4) Product Images from "DNA demethylation induces SALL4 gene re-expression in subgroups of hepatocellular carcinoma associated with Hepatitis B or C virus infection"

    Article Title: DNA demethylation induces SALL4 gene re-expression in subgroups of hepatocellular carcinoma associated with Hepatitis B or C virus infection

    Journal: Oncogene

    doi: 10.1038/onc.2016.399

    BRG1/BAF complex is involved in SALL4 re-expression in HBV replicating hepatocytes ( a, b ) PCR quantification of SALL4 mRNA (Left panel), and immunoblots of SALL4 (Right panel), following transfection of BRG1 siRNA (siBRG1) ( a ), and BRG1 plasmid ( b ), in HepAD38 cells grown with HBV replication by tetracycline removal for 10 days. Results are from three independent RNA isolations performed in identical triplicates. Error bars represent S.D. * P
    Figure Legend Snippet: BRG1/BAF complex is involved in SALL4 re-expression in HBV replicating hepatocytes ( a, b ) PCR quantification of SALL4 mRNA (Left panel), and immunoblots of SALL4 (Right panel), following transfection of BRG1 siRNA (siBRG1) ( a ), and BRG1 plasmid ( b ), in HepAD38 cells grown with HBV replication by tetracycline removal for 10 days. Results are from three independent RNA isolations performed in identical triplicates. Error bars represent S.D. * P

    Techniques Used: Expressing, Polymerase Chain Reaction, Western Blot, Transfection, Plasmid Preparation

    HBV replication enables binding and interaction of STAT3, OCT4 and BRG1 in SALL4 regulatory region ( a ) ChIP assays with STAT3, OCT4, and BRG1 antibodies, as indicated, employing HepAD38 cells grown −/+ HBV replication by tetracycline removal for D0 and D10, and SALL4 primers spanning CpG sites shown in Fig.1b . Data were quantified as % of input. ( b ) Co-immunoprecipitations (co-IPs) of STAT3, OCT4 and BRG1 using indicated antibodies, employing extracts from HepAD38 cells grown −/+ HBV replication by tetracycline removal for D0 and D10. A representative assay is shown from three independent experiments. ( c ) Sequential ChIP assays of HepAD38 cells −/+ HBV replication by tetracycline removal for D0 and D10, using BRG1 antibody followed by tandem IP with STAT3 or OCT4 antibody. Results are from three independent experiments. Data were quantified as % of input.
    Figure Legend Snippet: HBV replication enables binding and interaction of STAT3, OCT4 and BRG1 in SALL4 regulatory region ( a ) ChIP assays with STAT3, OCT4, and BRG1 antibodies, as indicated, employing HepAD38 cells grown −/+ HBV replication by tetracycline removal for D0 and D10, and SALL4 primers spanning CpG sites shown in Fig.1b . Data were quantified as % of input. ( b ) Co-immunoprecipitations (co-IPs) of STAT3, OCT4 and BRG1 using indicated antibodies, employing extracts from HepAD38 cells grown −/+ HBV replication by tetracycline removal for D0 and D10. A representative assay is shown from three independent experiments. ( c ) Sequential ChIP assays of HepAD38 cells −/+ HBV replication by tetracycline removal for D0 and D10, using BRG1 antibody followed by tandem IP with STAT3 or OCT4 antibody. Results are from three independent experiments. Data were quantified as % of input.

    Techniques Used: Binding Assay, Chromatin Immunoprecipitation

    5) Product Images from "The spring-loaded genome: Nucleosome redistributions are widespread, transient, and DNA-directed"

    Article Title: The spring-loaded genome: Nucleosome redistributions are widespread, transient, and DNA-directed

    Journal: Genome Research

    doi: 10.1101/gr.160150.113

    Nucleosome redistributions are determined by the underlying DNA sequence. ( A ) Western blots with the specified antibodies, at various times (in hours) after KSHV reactivation (hpr), of iSLK.219 cells treated with 0.2 µg/mL doxycycline. BRG1 protein
    Figure Legend Snippet: Nucleosome redistributions are determined by the underlying DNA sequence. ( A ) Western blots with the specified antibodies, at various times (in hours) after KSHV reactivation (hpr), of iSLK.219 cells treated with 0.2 µg/mL doxycycline. BRG1 protein

    Techniques Used: Sequencing, Western Blot

    6) Product Images from "The SWI/SNF subunits BRG1 affects alternative splicing by changing RNA binding factor interactions with RNA"

    Article Title: The SWI/SNF subunits BRG1 affects alternative splicing by changing RNA binding factor interactions with RNA

    Journal: bioRxiv

    doi: 10.1101/858852

    RNA binding factor are recruited by BRG1 and BRM to the target sites. A) ChIP-qPCR targeting MYL6 exon 6 in C33A cells expressing BRG1-wt and BRG1-mut was performed with antibodies against Sam68, hnRNPU, hnRNPL, DHX15, SYF1, THOC2, hnRNPA1 and hnRNP2B1; significant changes (p-value
    Figure Legend Snippet: RNA binding factor are recruited by BRG1 and BRM to the target sites. A) ChIP-qPCR targeting MYL6 exon 6 in C33A cells expressing BRG1-wt and BRG1-mut was performed with antibodies against Sam68, hnRNPU, hnRNPL, DHX15, SYF1, THOC2, hnRNPA1 and hnRNP2B1; significant changes (p-value

    Techniques Used: RNA Binding Assay, Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Expressing, Significance Assay

    Nucleosome and polymerase density in the affected exons A-C) ChIP-qPCR using antibodies against H3 (top panels) and H3K36me3 (bottom panels) targeting MYL6 gene (A), GADD45A exon 2 (B) and MAZ exon 5 (C) in C33A cells expressing BRG1 and BRG1-mut. H3K36me3 levels were normalized to H3. The association is related to the association in control cells, and significantly different values (p-value
    Figure Legend Snippet: Nucleosome and polymerase density in the affected exons A-C) ChIP-qPCR using antibodies against H3 (top panels) and H3K36me3 (bottom panels) targeting MYL6 gene (A), GADD45A exon 2 (B) and MAZ exon 5 (C) in C33A cells expressing BRG1 and BRG1-mut. H3K36me3 levels were normalized to H3. The association is related to the association in control cells, and significantly different values (p-value

    Techniques Used: Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Expressing, Significance Assay

    SWI/SNF interactions with RNA binding factor. A) Venn diagram with the number of RNPs interacting with BRG1 and BRM in the nascent RNA (data from Yu et al., 2018 ), that are associated with the GO term RNA splicing (GO:0008380). B) Number of BRG1 and BRM interacting proteins extracted from Yu et al. (2018) , known to be splicing (classification from Hegele et al., 2012 ). The total length of the bar corresponds to the total amount of proteins classified in each group. BRG1 interacting protein in blue, BRM in pink, both present in BRG1 and BRM in purple, and not present (NP) in any IP in grey. C) RNA binding proteins interacting with BRG1 or BRM ordered by their score from MASS-spectroscopy (left) with known motifs present in affected exons (right) classified into included (blue) or skipped (orange) exons. The RNA binding proteins selected for detailed mechanistic studies (hnRNPL, hnRNPU and Sam68) are depicted in bold-italics.
    Figure Legend Snippet: SWI/SNF interactions with RNA binding factor. A) Venn diagram with the number of RNPs interacting with BRG1 and BRM in the nascent RNA (data from Yu et al., 2018 ), that are associated with the GO term RNA splicing (GO:0008380). B) Number of BRG1 and BRM interacting proteins extracted from Yu et al. (2018) , known to be splicing (classification from Hegele et al., 2012 ). The total length of the bar corresponds to the total amount of proteins classified in each group. BRG1 interacting protein in blue, BRM in pink, both present in BRG1 and BRM in purple, and not present (NP) in any IP in grey. C) RNA binding proteins interacting with BRG1 or BRM ordered by their score from MASS-spectroscopy (left) with known motifs present in affected exons (right) classified into included (blue) or skipped (orange) exons. The RNA binding proteins selected for detailed mechanistic studies (hnRNPL, hnRNPU and Sam68) are depicted in bold-italics.

    Techniques Used: RNA Binding Assay, Mass Spectrometry

    BRG1 and BRM affect alternative splicing of a subset of genes. A) Venn diagram showing exons affected by the exogenous expression of BRG1-wt, BRG1-mut and BRM-wt. Exons affected only by BRG1-wt are referred to as “BRG1 ATPase dependent” while exons affected by both BRG1-wt and BRG1-mut are referred to as “BRG1 ATPase independent”. pOPRSVI plasmid was used as control. RNA was harvested 48 h after transfection and measured by RNA-seq, where differential exon levels were estimated with the MISO algorithm ( Katz et al., 2010 ). B) Number of exons with increased inclusion (blue) or skipped (orange) upon expression of BRG1. C) UCSC alternative events classification of BRG1 affected exons detected by MISO, represented as percentage of the total amount of affected exons detected. D) GC content in ATPase dependent (top panel) and ATPase independent (bottom panel) BRG1 affected exons +/- 500bp harbouring regions divided into included (blue) and skipped (orange) exons. Exons are plotted as 100 bp, each bp representing the average GC content of the 1% of the total length for each exon. Exons and the harbouring +/-500 bp regions show the 10% of GC in each position. E) Length of upstream and downstream introns from ATPase dependent (top panel) and ATPase independent (bottom panel) BRG1 affected exons. Bars represent the percentage of affected exons shorter or longer than 1 kb. F) Nucleosome density from ATPase dependent (top panel) and ATPase independent (bottom panel) BRG1 affected exons, generated with DeepTools2 (Ramírez et al., 2016) from publicly available data from the K562 cell line, at the exons differentially included (blue) or skipped (orange) by the SWI/SNF ATPases and the +/- 0.5 Kb harbouring region (light colour shows standard error).
    Figure Legend Snippet: BRG1 and BRM affect alternative splicing of a subset of genes. A) Venn diagram showing exons affected by the exogenous expression of BRG1-wt, BRG1-mut and BRM-wt. Exons affected only by BRG1-wt are referred to as “BRG1 ATPase dependent” while exons affected by both BRG1-wt and BRG1-mut are referred to as “BRG1 ATPase independent”. pOPRSVI plasmid was used as control. RNA was harvested 48 h after transfection and measured by RNA-seq, where differential exon levels were estimated with the MISO algorithm ( Katz et al., 2010 ). B) Number of exons with increased inclusion (blue) or skipped (orange) upon expression of BRG1. C) UCSC alternative events classification of BRG1 affected exons detected by MISO, represented as percentage of the total amount of affected exons detected. D) GC content in ATPase dependent (top panel) and ATPase independent (bottom panel) BRG1 affected exons +/- 500bp harbouring regions divided into included (blue) and skipped (orange) exons. Exons are plotted as 100 bp, each bp representing the average GC content of the 1% of the total length for each exon. Exons and the harbouring +/-500 bp regions show the 10% of GC in each position. E) Length of upstream and downstream introns from ATPase dependent (top panel) and ATPase independent (bottom panel) BRG1 affected exons. Bars represent the percentage of affected exons shorter or longer than 1 kb. F) Nucleosome density from ATPase dependent (top panel) and ATPase independent (bottom panel) BRG1 affected exons, generated with DeepTools2 (Ramírez et al., 2016) from publicly available data from the K562 cell line, at the exons differentially included (blue) or skipped (orange) by the SWI/SNF ATPases and the +/- 0.5 Kb harbouring region (light colour shows standard error).

    Techniques Used: Expressing, Plasmid Preparation, Transfection, RNA Sequencing Assay, Generated

    Model: BRG1 changes the recruitment pattern of RNA binding factor to alternative splicing exons in a context specific manner. MYL6 exon 6 has hnRNPU present in its vicinity at both chromatin and RNA level. Expression of BRG1 favours the recruitment of hnRNPL to this site in chromatin, while at the RNA level it recruits Sam68 and removes hnRNPU.
    Figure Legend Snippet: Model: BRG1 changes the recruitment pattern of RNA binding factor to alternative splicing exons in a context specific manner. MYL6 exon 6 has hnRNPU present in its vicinity at both chromatin and RNA level. Expression of BRG1 favours the recruitment of hnRNPL to this site in chromatin, while at the RNA level it recruits Sam68 and removes hnRNPU.

    Techniques Used: RNA Binding Assay, Expressing

    7) Product Images from "DNA demethylation induces SALL4 gene re-expression in subgroups of hepatocellular carcinoma associated with Hepatitis B or C virus infection"

    Article Title: DNA demethylation induces SALL4 gene re-expression in subgroups of hepatocellular carcinoma associated with Hepatitis B or C virus infection

    Journal: Oncogene

    doi: 10.1038/onc.2016.399

    BRG1/BAF complex is involved in SALL4 re-expression in HBV replicating hepatocytes
    Figure Legend Snippet: BRG1/BAF complex is involved in SALL4 re-expression in HBV replicating hepatocytes

    Techniques Used: Expressing

    HBV replication enables binding and interaction of STAT3, OCT4 and BRG1 in SALL4 regulatory region
    Figure Legend Snippet: HBV replication enables binding and interaction of STAT3, OCT4 and BRG1 in SALL4 regulatory region

    Techniques Used: Binding Assay

    Related Articles

    Immunohistochemistry:

    Article Title: Essential Roles of the Chromatin Remodeling Factor Brg1 in Spermatogenesis in Mice 1
    Article Snippet: .. Primary antibodies used were anti-EE2 (1:100; gift of Dr. Hiromitsu Tanaka), anti-BRG1 (1:100; 110641; Abcam), anti-BRM (1:100; 15597; Abcam), anti-H3K9me3 (1:100; 8898; Abcam), anti-phospho-S5 Pol II (1:100; IHC-00387; Bethyl Laboratories), anti-cH2AX (1:100; 05–636; Millipore), anti-RAD51 (1:100; 63801; Abcam), and anti-H1t (1:400; gift of Dr. Mary Ann Handel). .. Apoptosis was analyzed on cryosections using the ApopTag Fluorescein In Situ Apoptosis Detection Kit (Chemicon International).

    Article Title: SMARCA4 loss is synthetic lethal with CDK4/6 inhibition in non-small cell lung cancer
    Article Snippet: .. IHC was performed on the TMAs using anti-SMARCA4 (anti-BRG1 antibody (clone EPNCIR111A, dilution, 1:100 dilution, Abcam, Cambridge, UK), anti-p16 (clone G175-405, 1:20 dilution, BD PharmingenTM ) and anti-RB1 (G3-245, 1:100 dilution, BD PharmingenTM ) antibodies. .. For assessment of SMARCA4, SMARCA2 and RB1, unequivocally absent staining in the nuclei of viable tumor cells as opposed to strong staining in background stromal cells was considered IHC negative.

    Cell Culture:

    Article Title: The SWI/SNF Chromatin Regulator BRG1 Modulates the Transcriptional Regulatory Activity of the Epstein-Barr Virus DNA Polymerase Processivity Factor BMRF1
    Article Snippet: .. Briefly, the cultured slides were fixed with 4% paraformaldehyde and incubated with anti-Flag (M2; Sigma-Aldrich), anti-HA (HA.11; Covance), or anti-BRG1 (EPNCIR111A; Abcam) at 4°C overnight. .. Following three washes with PBS, the slides were incubated with a fluorescein isothiocyanate (FITC)- or a rhodamine-conjugated secondary antibody at 37°C for 1 h. Finally, the slides were stained with 100 ng/ml Hoechst 33258 (Sigma-Aldrich) for 5 min at room temperature.

    Immunoprecipitation:

    Article Title: Transcriptional regulation of Munc13-4 expression in cytotoxic lymphocytes is disrupted by an intronic mutation associated with a primary immunodeficiency
    Article Snippet: .. For each sample, 106 cell equivalents of chromatin were immunoprecipitated with the EZ-ChIP chromatin immunoprecipitation kit (Millipore) using 1 µg each of the following ChIP-grade antibodies: mouse IgG (Millipore), RNA polymerase II (Millipore), STAT4 (Abcam), anti-histone H3 (acetyl K27; Abcam), anti-BRG1 (Abcam), and anti-histone H3 (tri-methyl K4; Abcam). .. Cross-links were reversed by incubation at 65°C with the addition of proteinase K overnight.

    Incubation:

    Article Title: BRG1 in the nucleus accumbens regulates cocaine-seeking behavior
    Article Snippet: .. Membranes were incubated overnight at 4°C with primary antibodies diluted in blocking buffer (Rockland Immunochemicals, Inc., Limerick, PA), including: rabbit anti-BRG1 (1:2,000; Abcam, Cambridge, MA), rabbit anti-phospho(p)-SMAD3 (1:500; Calbiochem of Millipore Corp., Billerica, MA), and mouse anti-β-actin (1:10,000; Cell Signaling Technologies, Inc., Danvers, MA). .. Membranes were incubated with IRDye secondary antibodies (1:5,000; LI-COR, Inc., Lincoln, NE) for 1 h at room temperature.

    Article Title: The SWI/SNF Chromatin Regulator BRG1 Modulates the Transcriptional Regulatory Activity of the Epstein-Barr Virus DNA Polymerase Processivity Factor BMRF1
    Article Snippet: .. Briefly, the cultured slides were fixed with 4% paraformaldehyde and incubated with anti-Flag (M2; Sigma-Aldrich), anti-HA (HA.11; Covance), or anti-BRG1 (EPNCIR111A; Abcam) at 4°C overnight. .. Following three washes with PBS, the slides were incubated with a fluorescein isothiocyanate (FITC)- or a rhodamine-conjugated secondary antibody at 37°C for 1 h. Finally, the slides were stained with 100 ng/ml Hoechst 33258 (Sigma-Aldrich) for 5 min at room temperature.

    Blocking Assay:

    Article Title: BRG1 in the nucleus accumbens regulates cocaine-seeking behavior
    Article Snippet: .. Membranes were incubated overnight at 4°C with primary antibodies diluted in blocking buffer (Rockland Immunochemicals, Inc., Limerick, PA), including: rabbit anti-BRG1 (1:2,000; Abcam, Cambridge, MA), rabbit anti-phospho(p)-SMAD3 (1:500; Calbiochem of Millipore Corp., Billerica, MA), and mouse anti-β-actin (1:10,000; Cell Signaling Technologies, Inc., Danvers, MA). .. Membranes were incubated with IRDye secondary antibodies (1:5,000; LI-COR, Inc., Lincoln, NE) for 1 h at room temperature.

    Chromatin Immunoprecipitation:

    Article Title: Transcriptional regulation of Munc13-4 expression in cytotoxic lymphocytes is disrupted by an intronic mutation associated with a primary immunodeficiency
    Article Snippet: .. For each sample, 106 cell equivalents of chromatin were immunoprecipitated with the EZ-ChIP chromatin immunoprecipitation kit (Millipore) using 1 µg each of the following ChIP-grade antibodies: mouse IgG (Millipore), RNA polymerase II (Millipore), STAT4 (Abcam), anti-histone H3 (acetyl K27; Abcam), anti-BRG1 (Abcam), and anti-histone H3 (tri-methyl K4; Abcam). .. Cross-links were reversed by incubation at 65°C with the addition of proteinase K overnight.

    Article Title: Long non-coding RNA urothelial carcinoma associated 1 induces cell replication by inhibiting BRG1 in 5637 cells
    Article Snippet: .. ChIP was performed overnight at 4°C using the BRG1 antibody (ab4081; Abcam) or an isotype control IgG. .. After a 1 h incubation in the presence of salmon sperm DNA/protein A agarose beads, the immunoprecipitated DNA/protein complexes were then washed and eluted from the beads with 1% SDS and 0.1 M NaHCO3 solution.

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  • 96
    Abcam anti smarca4
    <t>SMARCA4/2</t> regulate CCND1 via controlling chromatin accessibility and upregulating JUN . a Assay for transposase-accessible chromatin sequencing (ATAC-seq) and chromatin immunoprecipitation sequencing (ChIP-seq) data in vicinity of the CCND1 locus indicate enhanced chromatin accessibility upon SMARCA4/2 restoration. Note SMARCA4 at CCND1 promoter and formation of new putative enhancer ~50 kb upstream of CCND1 . Track height is normalized to relative number of mapped reads. b Zoomed-in view of the putative CCND1 enhancer region. Shown are ATAC-seq peaks in H1703 cells before and after SMARCA4/2 restoration and the publicly available c-Fos/c-Jun ChIP data of endothelial cell line, human umbilical vein endothelial cell (HUVEC) (GSM935585, GSM935278). Location of canonical adaptor protein-1 (AP-1) motifs are indicated. c ATAC and ChIP-seq data in vicinity of JUN locus as described in a . Note SMARCA4 at JUN promoter and extensive opening of nearby putative enhancers. d– i Restoration of SMARCA4 in H1703 ( d , e ) and H1299 ( f , g ) or SMARCA2 restoration in H1703 ( h , i ) cells upregulate c-Jun messenger RNA (mRNA) ( d , f , h ) and protein ( e , g , i ). j , k Knockdown of JUN partially abrogated SMARCA4-mediated induction of cyclin D1 mRNA ( j ) and protein ( k ) expression in H1703 cells. l Proposed model showing that SMARCA4 directly regulates CCND1 and also upregulates JUN which positively regulates CCND1 . Two-tailed t -test. Error bars represent mean ± s.d., *** p
    Anti Smarca4, supplied by Abcam, used in various techniques. Bioz Stars score: 96/100, based on 25 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    94
    Abcam anti brg1 antibody
    <t>BRG1</t> and miR-148b are expressed at low levels in lung cancer tissues and cell lines. Notes: ( A ) BRG1 mRNA level is lower in lung cancer tissues than that in adjacent normal lung tissues. ( B ) The expression level of BRG1 protein is decreased in lung cancer tissues compared to that in adjacent normal lung tissues. ( C ) BRG1 mRNA levels in lung cancer cell lines A549, H522, and H1299 are lower than those in lung cell lines WI38, MRC5, and Beas-2B. ( D ) BRG1 protein levels in lung cancer cell lines A549, H522, and H1299 are lower than those in lung cell lines WI38, MRC5, and Beas-2B. ( E and F ) miR-148b expression level is significantly decreased in lung cancer tissues and cells. * P
    Anti Brg1 Antibody, supplied by Abcam, used in various techniques. Bioz Stars score: 94/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    99
    Abcam anti brg1
    Effect of SWI/SNF subunits BAF47, BAF53a and <t>BRG1</t> knockdown on the irreversible cell cycle exit. C2C12 myoblasts were transfected with control, BRG1, BAF53a or BAF47 siRNAs and cells were labeled with BrdU either after 96 h in DM or 12 h after the switch back to GM (96+12 h GM). We show one representative of 3 independent experiments. Left panel: BrdU indexes ratio of BrdU positive nuclei/DAPI labeled nuclei) are shown in the histogram, error bars represent SEM from 3 quantifications of the presented experiment (*p
    Anti Brg1, supplied by Abcam, used in various techniques. Bioz Stars score: 99/100, based on 6 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    SMARCA4/2 regulate CCND1 via controlling chromatin accessibility and upregulating JUN . a Assay for transposase-accessible chromatin sequencing (ATAC-seq) and chromatin immunoprecipitation sequencing (ChIP-seq) data in vicinity of the CCND1 locus indicate enhanced chromatin accessibility upon SMARCA4/2 restoration. Note SMARCA4 at CCND1 promoter and formation of new putative enhancer ~50 kb upstream of CCND1 . Track height is normalized to relative number of mapped reads. b Zoomed-in view of the putative CCND1 enhancer region. Shown are ATAC-seq peaks in H1703 cells before and after SMARCA4/2 restoration and the publicly available c-Fos/c-Jun ChIP data of endothelial cell line, human umbilical vein endothelial cell (HUVEC) (GSM935585, GSM935278). Location of canonical adaptor protein-1 (AP-1) motifs are indicated. c ATAC and ChIP-seq data in vicinity of JUN locus as described in a . Note SMARCA4 at JUN promoter and extensive opening of nearby putative enhancers. d– i Restoration of SMARCA4 in H1703 ( d , e ) and H1299 ( f , g ) or SMARCA2 restoration in H1703 ( h , i ) cells upregulate c-Jun messenger RNA (mRNA) ( d , f , h ) and protein ( e , g , i ). j , k Knockdown of JUN partially abrogated SMARCA4-mediated induction of cyclin D1 mRNA ( j ) and protein ( k ) expression in H1703 cells. l Proposed model showing that SMARCA4 directly regulates CCND1 and also upregulates JUN which positively regulates CCND1 . Two-tailed t -test. Error bars represent mean ± s.d., *** p

    Journal: Nature Communications

    Article Title: SMARCA4 loss is synthetic lethal with CDK4/6 inhibition in non-small cell lung cancer

    doi: 10.1038/s41467-019-08380-1

    Figure Lengend Snippet: SMARCA4/2 regulate CCND1 via controlling chromatin accessibility and upregulating JUN . a Assay for transposase-accessible chromatin sequencing (ATAC-seq) and chromatin immunoprecipitation sequencing (ChIP-seq) data in vicinity of the CCND1 locus indicate enhanced chromatin accessibility upon SMARCA4/2 restoration. Note SMARCA4 at CCND1 promoter and formation of new putative enhancer ~50 kb upstream of CCND1 . Track height is normalized to relative number of mapped reads. b Zoomed-in view of the putative CCND1 enhancer region. Shown are ATAC-seq peaks in H1703 cells before and after SMARCA4/2 restoration and the publicly available c-Fos/c-Jun ChIP data of endothelial cell line, human umbilical vein endothelial cell (HUVEC) (GSM935585, GSM935278). Location of canonical adaptor protein-1 (AP-1) motifs are indicated. c ATAC and ChIP-seq data in vicinity of JUN locus as described in a . Note SMARCA4 at JUN promoter and extensive opening of nearby putative enhancers. d– i Restoration of SMARCA4 in H1703 ( d , e ) and H1299 ( f , g ) or SMARCA2 restoration in H1703 ( h , i ) cells upregulate c-Jun messenger RNA (mRNA) ( d , f , h ) and protein ( e , g , i ). j , k Knockdown of JUN partially abrogated SMARCA4-mediated induction of cyclin D1 mRNA ( j ) and protein ( k ) expression in H1703 cells. l Proposed model showing that SMARCA4 directly regulates CCND1 and also upregulates JUN which positively regulates CCND1 . Two-tailed t -test. Error bars represent mean ± s.d., *** p

    Article Snippet: IHC was performed on the TMAs using anti-SMARCA4 (anti-BRG1 antibody (clone EPNCIR111A, dilution, 1:100 dilution, Abcam, Cambridge, UK), anti-p16 (clone G175-405, 1:20 dilution, BD PharmingenTM ) and anti-RB1 (G3-245, 1:100 dilution, BD PharmingenTM ) antibodies.

    Techniques: Sequencing, ChIP-sequencing, Chromatin Immunoprecipitation, Expressing, Two Tailed Test

    SMARCA4 loss is synthetic lethal with cyclin-dependent kinase 4/6 (CDK4/6) inhibition in non-small cell lung cancer (NSCLC). a , b SMARCA4 restoration in SMARCA4-deficient cell lines confers drug resistance to palbociclib. Colony formation assays of H1299 ( a ) and H1703 ( b ) cells expressing vector control or SMARCA4 and treated with palbociclib (H1299, 300 nM; H1703, 100 nM). c SMARCA2 knockdown in SMARCA4-deficient H1299 cells sensitizes cells to palbociclib treatment. Colony formation assay of H1299 cells expressing pLKO control or SMARCA2 short hairpin RNAs (shRNAs) and treated with palbociclib. d SMARCA2 restoration in SMARCA4/2-dual deficient cells H1703 confers resistance to palbociclib. Colony formation assay of H1703 cells expressing vector control or SMARCA2 and treated with palbociclib. e – g Resistance to palbociclib after restoration of SMARCA4 is also observed in mouse xenograft models using an isogenic cell pair of H1299 cells expressing vector control or SMARCA4 . e Tumor volume evolution during the course of the experiment in H1299 xenograft models expressing vector control (left) or SMARCA4 (right). f Tumor volume fold change during the establishment phase (left) and during palbociclib treatment (right) in the same models. g Immunohistochemistry (IHC) analysis of SMARCA4 in the representative endpoint tumors of H1703 control or SMARCA4-restored from above. Bar 50 µm. Error bars represent mean ± standard error of mean (s.e.m.); two-way analysis of variance (ANOVA), **** p

    Journal: Nature Communications

    Article Title: SMARCA4 loss is synthetic lethal with CDK4/6 inhibition in non-small cell lung cancer

    doi: 10.1038/s41467-019-08380-1

    Figure Lengend Snippet: SMARCA4 loss is synthetic lethal with cyclin-dependent kinase 4/6 (CDK4/6) inhibition in non-small cell lung cancer (NSCLC). a , b SMARCA4 restoration in SMARCA4-deficient cell lines confers drug resistance to palbociclib. Colony formation assays of H1299 ( a ) and H1703 ( b ) cells expressing vector control or SMARCA4 and treated with palbociclib (H1299, 300 nM; H1703, 100 nM). c SMARCA2 knockdown in SMARCA4-deficient H1299 cells sensitizes cells to palbociclib treatment. Colony formation assay of H1299 cells expressing pLKO control or SMARCA2 short hairpin RNAs (shRNAs) and treated with palbociclib. d SMARCA2 restoration in SMARCA4/2-dual deficient cells H1703 confers resistance to palbociclib. Colony formation assay of H1703 cells expressing vector control or SMARCA2 and treated with palbociclib. e – g Resistance to palbociclib after restoration of SMARCA4 is also observed in mouse xenograft models using an isogenic cell pair of H1299 cells expressing vector control or SMARCA4 . e Tumor volume evolution during the course of the experiment in H1299 xenograft models expressing vector control (left) or SMARCA4 (right). f Tumor volume fold change during the establishment phase (left) and during palbociclib treatment (right) in the same models. g Immunohistochemistry (IHC) analysis of SMARCA4 in the representative endpoint tumors of H1703 control or SMARCA4-restored from above. Bar 50 µm. Error bars represent mean ± standard error of mean (s.e.m.); two-way analysis of variance (ANOVA), **** p

    Article Snippet: IHC was performed on the TMAs using anti-SMARCA4 (anti-BRG1 antibody (clone EPNCIR111A, dilution, 1:100 dilution, Abcam, Cambridge, UK), anti-p16 (clone G175-405, 1:20 dilution, BD PharmingenTM ) and anti-RB1 (G3-245, 1:100 dilution, BD PharmingenTM ) antibodies.

    Techniques: Inhibition, Expressing, Plasmid Preparation, Colony Assay, Immunohistochemistry

    Reduced cyclin D1 in SMARCA4-deficient non-small cell lung cancer (NSCLC) cells causessensitivities to cyclin-dependent kinase 4/6 (CDK4/6) inhibitors. a , b SMARCA4-deficient NSCLC cell lines express reduced cyclin D1 levels. Western blot analysis for the indicated proteins ( a ) and CCND1 messenger RNA (mRNA) expression ( b ) of a panel of NSCLC cell lines. HSP90 was used as a loading control. Relative CCND1 mRNA expression (relative to GAPDH ) was measured by real-time quantitative reverse transcription PCR (RT-qPCR). A4: SMARCA4, A4/2: SMARCA4/2, Pro: proficient, Def: deficient, K: KRAS mutation. Empty triangles indicate RB-deficient cell lines. Turquoise color indicates cell lines with KRAS mutation. Error bars: mean ± standard deviation (s.d.) of biological replicates ( n = 3); two-tailed t test, * p

    Journal: Nature Communications

    Article Title: SMARCA4 loss is synthetic lethal with CDK4/6 inhibition in non-small cell lung cancer

    doi: 10.1038/s41467-019-08380-1

    Figure Lengend Snippet: Reduced cyclin D1 in SMARCA4-deficient non-small cell lung cancer (NSCLC) cells causessensitivities to cyclin-dependent kinase 4/6 (CDK4/6) inhibitors. a , b SMARCA4-deficient NSCLC cell lines express reduced cyclin D1 levels. Western blot analysis for the indicated proteins ( a ) and CCND1 messenger RNA (mRNA) expression ( b ) of a panel of NSCLC cell lines. HSP90 was used as a loading control. Relative CCND1 mRNA expression (relative to GAPDH ) was measured by real-time quantitative reverse transcription PCR (RT-qPCR). A4: SMARCA4, A4/2: SMARCA4/2, Pro: proficient, Def: deficient, K: KRAS mutation. Empty triangles indicate RB-deficient cell lines. Turquoise color indicates cell lines with KRAS mutation. Error bars: mean ± standard deviation (s.d.) of biological replicates ( n = 3); two-tailed t test, * p

    Article Snippet: IHC was performed on the TMAs using anti-SMARCA4 (anti-BRG1 antibody (clone EPNCIR111A, dilution, 1:100 dilution, Abcam, Cambridge, UK), anti-p16 (clone G175-405, 1:20 dilution, BD PharmingenTM ) and anti-RB1 (G3-245, 1:100 dilution, BD PharmingenTM ) antibodies.

    Techniques: Western Blot, Expressing, Polymerase Chain Reaction, Quantitative RT-PCR, Mutagenesis, Standard Deviation, Two Tailed Test

    Palbociclib is effective against SMARCA4-deficient non-small cell lung cancer (NSCLC) tumor growth in vivo. Palbociclib inhibits tumor growth in xenograft models of H1299 ( a , b , e , f ) and H1703 ( c , d , g , h ). a , c Tumor size from day 0 of treatment in H1299 ( a , n = 4 per group) and H1703 ( c , n = 8 for vehicle, n = 7 for palbociclib; 150 mg kg −1 ) models. Error bars represent mean ± standard error of mean (s.e.m.); two-way analysis of variance (ANOVA), **** p

    Journal: Nature Communications

    Article Title: SMARCA4 loss is synthetic lethal with CDK4/6 inhibition in non-small cell lung cancer

    doi: 10.1038/s41467-019-08380-1

    Figure Lengend Snippet: Palbociclib is effective against SMARCA4-deficient non-small cell lung cancer (NSCLC) tumor growth in vivo. Palbociclib inhibits tumor growth in xenograft models of H1299 ( a , b , e , f ) and H1703 ( c , d , g , h ). a , c Tumor size from day 0 of treatment in H1299 ( a , n = 4 per group) and H1703 ( c , n = 8 for vehicle, n = 7 for palbociclib; 150 mg kg −1 ) models. Error bars represent mean ± standard error of mean (s.e.m.); two-way analysis of variance (ANOVA), **** p

    Article Snippet: IHC was performed on the TMAs using anti-SMARCA4 (anti-BRG1 antibody (clone EPNCIR111A, dilution, 1:100 dilution, Abcam, Cambridge, UK), anti-p16 (clone G175-405, 1:20 dilution, BD PharmingenTM ) and anti-RB1 (G3-245, 1:100 dilution, BD PharmingenTM ) antibodies.

    Techniques: In Vivo

    Extensive opening of regulatory elements by induction of SMARCA4/2. a Venn diagram showing overlap of open chromatin sites in control infected H1703 cells with or without SMARCA4 overexpression. Note the dramatic increase in open chromatin sites upon SMARCA4 overexpression. b Distribution of SMARCA4-dependent and -independent open chromatin sites relative to nearest gene transcriptional start site. Note that SMARCA4-dependent sites are much less likely to be promoters (within 1 kb of transcription start site (TSS)). c , d Metaplot ( c ) and heatmap ( d ) of assay for transposase-accessible chromatin sequencing (ATAC-seq) read data from control-infected, SMARCA4-infected and SMARCA2-infected cells over the 62,878 SMARCA4-dependent ATAC peaks. Note similar effect of SMARCA2 and SMARCA4. e , f Metaplot ( e ) and heatmap ( f ) of H3K27Ac chromatin immunoprecipitation (ChIP) data from control-transfected, SMARCA4-infected and SMARCA2-infected cells over the 62,878 SMARCA4-dependent ATAC peaks

    Journal: Nature Communications

    Article Title: SMARCA4 loss is synthetic lethal with CDK4/6 inhibition in non-small cell lung cancer

    doi: 10.1038/s41467-019-08380-1

    Figure Lengend Snippet: Extensive opening of regulatory elements by induction of SMARCA4/2. a Venn diagram showing overlap of open chromatin sites in control infected H1703 cells with or without SMARCA4 overexpression. Note the dramatic increase in open chromatin sites upon SMARCA4 overexpression. b Distribution of SMARCA4-dependent and -independent open chromatin sites relative to nearest gene transcriptional start site. Note that SMARCA4-dependent sites are much less likely to be promoters (within 1 kb of transcription start site (TSS)). c , d Metaplot ( c ) and heatmap ( d ) of assay for transposase-accessible chromatin sequencing (ATAC-seq) read data from control-infected, SMARCA4-infected and SMARCA2-infected cells over the 62,878 SMARCA4-dependent ATAC peaks. Note similar effect of SMARCA2 and SMARCA4. e , f Metaplot ( e ) and heatmap ( f ) of H3K27Ac chromatin immunoprecipitation (ChIP) data from control-transfected, SMARCA4-infected and SMARCA2-infected cells over the 62,878 SMARCA4-dependent ATAC peaks

    Article Snippet: IHC was performed on the TMAs using anti-SMARCA4 (anti-BRG1 antibody (clone EPNCIR111A, dilution, 1:100 dilution, Abcam, Cambridge, UK), anti-p16 (clone G175-405, 1:20 dilution, BD PharmingenTM ) and anti-RB1 (G3-245, 1:100 dilution, BD PharmingenTM ) antibodies.

    Techniques: Infection, Over Expression, Sequencing, Chromatin Immunoprecipitation, Transfection

    SMARCA4/2 loss causes reduced cyclin D1 expression in non-small cell lung cancer (NSCLC). a – d SMARCA4/2 regulate cyclin D1 expression in NSCLC. a , b SMARCA4 restoration upregulates cyclin D1 protein (left) and messenger RNA (mRNA) (right) expression in H1299 ( a ) and H1703 ( b ) cells. c SMARCA2 knockdown in H1299 cells suppresses cyclin D1 protein (left) and mRNA (right) expression. d SMARCA2 restoration in H1703 cells elevates cyclin D1 protein (left) and mRNA (right) expression. Relative CCND1 mRNA expression (relative to GAPDH ) was measured by real-time quantitative reverse transcription PCR (RT-qPCR). Error bars: mean ± s.d. of biological replicates ( n = 3, two-tailed t -test, * p

    Journal: Nature Communications

    Article Title: SMARCA4 loss is synthetic lethal with CDK4/6 inhibition in non-small cell lung cancer

    doi: 10.1038/s41467-019-08380-1

    Figure Lengend Snippet: SMARCA4/2 loss causes reduced cyclin D1 expression in non-small cell lung cancer (NSCLC). a – d SMARCA4/2 regulate cyclin D1 expression in NSCLC. a , b SMARCA4 restoration upregulates cyclin D1 protein (left) and messenger RNA (mRNA) (right) expression in H1299 ( a ) and H1703 ( b ) cells. c SMARCA2 knockdown in H1299 cells suppresses cyclin D1 protein (left) and mRNA (right) expression. d SMARCA2 restoration in H1703 cells elevates cyclin D1 protein (left) and mRNA (right) expression. Relative CCND1 mRNA expression (relative to GAPDH ) was measured by real-time quantitative reverse transcription PCR (RT-qPCR). Error bars: mean ± s.d. of biological replicates ( n = 3, two-tailed t -test, * p

    Article Snippet: IHC was performed on the TMAs using anti-SMARCA4 (anti-BRG1 antibody (clone EPNCIR111A, dilution, 1:100 dilution, Abcam, Cambridge, UK), anti-p16 (clone G175-405, 1:20 dilution, BD PharmingenTM ) and anti-RB1 (G3-245, 1:100 dilution, BD PharmingenTM ) antibodies.

    Techniques: Expressing, Polymerase Chain Reaction, Quantitative RT-PCR, Two Tailed Test

    BRG1 is required for hSNF5-mediated induction of p15 INK4b and p16 INK4a . (A) Western blot analysis of the BRG1 protein levels in MON cell extracts prepared 4 days after BRG1 knockdown using lentiviral transduction with viruses expressing shRNA targeting BRG1 mRNA (clone 15549; Open Biosystems; top panel, lanes 3 and 4). As a control, cells were transduced with lentiviruses expressing GFP. One day after transduction with either GFP- or shRNA-expressing lentiviruses, cells were transduced again with viruses expressing either GFP (middle panel, lanes 1 and 3) or hSNF5 (lanes 2 and 3). Cell extracts were prepared 72 h after hSNF5 expression. Histone H3 serves as a loading control. (B) Loss of BRG1 abrogates transcriptional activation of p15 INK4b and p16 INK4a by hSNF5. Relative expression levels of p15 INK4b , p14 ARF , and p16 INK4a in these cells were determined by RT-qPCR of isolated mRNA, 72 h after hSNF5 expression. The bar graphs represent the mean of three independent experiments, each analyzed in triplicate by RT-qPCR.

    Journal: Molecular and Cellular Biology

    Article Title: SWI/SNF Mediates Polycomb Eviction and Epigenetic Reprogramming of the INK4b-ARF-INK4a Locus

    doi: 10.1128/MCB.02019-07

    Figure Lengend Snippet: BRG1 is required for hSNF5-mediated induction of p15 INK4b and p16 INK4a . (A) Western blot analysis of the BRG1 protein levels in MON cell extracts prepared 4 days after BRG1 knockdown using lentiviral transduction with viruses expressing shRNA targeting BRG1 mRNA (clone 15549; Open Biosystems; top panel, lanes 3 and 4). As a control, cells were transduced with lentiviruses expressing GFP. One day after transduction with either GFP- or shRNA-expressing lentiviruses, cells were transduced again with viruses expressing either GFP (middle panel, lanes 1 and 3) or hSNF5 (lanes 2 and 3). Cell extracts were prepared 72 h after hSNF5 expression. Histone H3 serves as a loading control. (B) Loss of BRG1 abrogates transcriptional activation of p15 INK4b and p16 INK4a by hSNF5. Relative expression levels of p15 INK4b , p14 ARF , and p16 INK4a in these cells were determined by RT-qPCR of isolated mRNA, 72 h after hSNF5 expression. The bar graphs represent the mean of three independent experiments, each analyzed in triplicate by RT-qPCR.

    Article Snippet: The following antibodies were used: SNF5 (ab12167 [Abcam]), BRG1 (ab4081 [Abcam]), SUZ12 (ab12073 [Abcam]), BMI1 (ab14389 [Abcam]), EZH2 (Sc-25383 [Santa Cruz]), DNMT3B (IMG-184A [IMGENEX]), POL II (Sc-899 [Santa Cruz]), mixed lineage leukemia 1 (MLL1; A300-086A [Bethyl Laboratories]), Histone H3 (ab1791 [Abcam]), H3-K4me3 (ab12209 [Abcam]), and H3-K27me3 (catalog no. 07-449 [Upstate]).

    Techniques: Western Blot, Transduction, Expressing, shRNA, Activation Assay, Quantitative RT-PCR, Isolation

    BRG1 and miR-148b are expressed at low levels in lung cancer tissues and cell lines. Notes: ( A ) BRG1 mRNA level is lower in lung cancer tissues than that in adjacent normal lung tissues. ( B ) The expression level of BRG1 protein is decreased in lung cancer tissues compared to that in adjacent normal lung tissues. ( C ) BRG1 mRNA levels in lung cancer cell lines A549, H522, and H1299 are lower than those in lung cell lines WI38, MRC5, and Beas-2B. ( D ) BRG1 protein levels in lung cancer cell lines A549, H522, and H1299 are lower than those in lung cell lines WI38, MRC5, and Beas-2B. ( E and F ) miR-148b expression level is significantly decreased in lung cancer tissues and cells. * P

    Journal: OncoTargets and therapy

    Article Title: Restoration of BRG1 inhibits proliferation and metastasis of lung cancer by regulating tumor suppressor miR-148b

    doi: 10.2147/OTT.S95500

    Figure Lengend Snippet: BRG1 and miR-148b are expressed at low levels in lung cancer tissues and cell lines. Notes: ( A ) BRG1 mRNA level is lower in lung cancer tissues than that in adjacent normal lung tissues. ( B ) The expression level of BRG1 protein is decreased in lung cancer tissues compared to that in adjacent normal lung tissues. ( C ) BRG1 mRNA levels in lung cancer cell lines A549, H522, and H1299 are lower than those in lung cell lines WI38, MRC5, and Beas-2B. ( D ) BRG1 protein levels in lung cancer cell lines A549, H522, and H1299 are lower than those in lung cell lines WI38, MRC5, and Beas-2B. ( E and F ) miR-148b expression level is significantly decreased in lung cancer tissues and cells. * P

    Article Snippet: The sheared chromatin was then immunoprecipitated by using anti-BRG1 antibody (Abcam, Cambridge, UK) or control IgG (Sigma, St. Louis, MO, USA).

    Techniques: Expressing

    Restoration of BRG1 inhibits proliferation and metastasis of lung cancer cells but downregulation of miR-148b resists the effects. Notes: ( A and B ) Restoration of BRG1 inhibits proliferation of A549 and H522 cells, respectively. Downregulation of miR-148b shows an opposite effect. ( C and D ) Overexpression of BRG1 reduces the expression levels of Ki67 in A549 and H522 cells, respectively, but downregulation of miR-148b elevates them. ( E and F ) Restoration of BRG1 suppresses the migration ability of A549 and H522 cells, respectively, which is resisted by downregulation of miR-148b. ( G and H ) TGFβ1 decreases the level of E-cadherin but increases the levels of N-cadherin and vimentin, enhancing EMT of A549 and H522 cell lines, respectively. Restoration of BRG1 suppresses TGFβ1-induced EMT, but downregulation of miR-148b reverses it. * P

    Journal: OncoTargets and therapy

    Article Title: Restoration of BRG1 inhibits proliferation and metastasis of lung cancer by regulating tumor suppressor miR-148b

    doi: 10.2147/OTT.S95500

    Figure Lengend Snippet: Restoration of BRG1 inhibits proliferation and metastasis of lung cancer cells but downregulation of miR-148b resists the effects. Notes: ( A and B ) Restoration of BRG1 inhibits proliferation of A549 and H522 cells, respectively. Downregulation of miR-148b shows an opposite effect. ( C and D ) Overexpression of BRG1 reduces the expression levels of Ki67 in A549 and H522 cells, respectively, but downregulation of miR-148b elevates them. ( E and F ) Restoration of BRG1 suppresses the migration ability of A549 and H522 cells, respectively, which is resisted by downregulation of miR-148b. ( G and H ) TGFβ1 decreases the level of E-cadherin but increases the levels of N-cadherin and vimentin, enhancing EMT of A549 and H522 cell lines, respectively. Restoration of BRG1 suppresses TGFβ1-induced EMT, but downregulation of miR-148b reverses it. * P

    Article Snippet: The sheared chromatin was then immunoprecipitated by using anti-BRG1 antibody (Abcam, Cambridge, UK) or control IgG (Sigma, St. Louis, MO, USA).

    Techniques: Over Expression, Expressing, Migration

    BRG1 binds to miR-148b target gene promoter in lung cancer cells. Notes: ( A ) The change of BRG1 levels after treatment with pcDNA-BRG1 and si-BRG1 in A549 cells. ( B ) miR-148b levels are positively correlated with BRG1 expression levels in A549 cells. ( C ) The change of BRG1 levels after treatment with pcDNA-BRG1 and si-BRG1 in H522 cells. ( D ) miR-148b levels are positively correlated with BRG1 expression levels in H522 cells. ( E ) Online platform ChIPBase displays the integrated TF-miRNAs network of miR-148b. ( F ) ChIP assay shows that BRG1 binds to the promoter of miR-148b target gene in A549 cells. * P

    Journal: OncoTargets and therapy

    Article Title: Restoration of BRG1 inhibits proliferation and metastasis of lung cancer by regulating tumor suppressor miR-148b

    doi: 10.2147/OTT.S95500

    Figure Lengend Snippet: BRG1 binds to miR-148b target gene promoter in lung cancer cells. Notes: ( A ) The change of BRG1 levels after treatment with pcDNA-BRG1 and si-BRG1 in A549 cells. ( B ) miR-148b levels are positively correlated with BRG1 expression levels in A549 cells. ( C ) The change of BRG1 levels after treatment with pcDNA-BRG1 and si-BRG1 in H522 cells. ( D ) miR-148b levels are positively correlated with BRG1 expression levels in H522 cells. ( E ) Online platform ChIPBase displays the integrated TF-miRNAs network of miR-148b. ( F ) ChIP assay shows that BRG1 binds to the promoter of miR-148b target gene in A549 cells. * P

    Article Snippet: The sheared chromatin was then immunoprecipitated by using anti-BRG1 antibody (Abcam, Cambridge, UK) or control IgG (Sigma, St. Louis, MO, USA).

    Techniques: Expressing, Chromatin Immunoprecipitation

    Effect of SWI/SNF subunits BAF47, BAF53a and BRG1 knockdown on the irreversible cell cycle exit. C2C12 myoblasts were transfected with control, BRG1, BAF53a or BAF47 siRNAs and cells were labeled with BrdU either after 96 h in DM or 12 h after the switch back to GM (96+12 h GM). We show one representative of 3 independent experiments. Left panel: BrdU indexes ratio of BrdU positive nuclei/DAPI labeled nuclei) are shown in the histogram, error bars represent SEM from 3 quantifications of the presented experiment (*p

    Journal: PLoS ONE

    Article Title: The SWI/SNF Subunit/Tumor Suppressor BAF47/INI1 Is Essential in Cell Cycle Arrest upon Skeletal Muscle Terminal Differentiation

    doi: 10.1371/journal.pone.0108858

    Figure Lengend Snippet: Effect of SWI/SNF subunits BAF47, BAF53a and BRG1 knockdown on the irreversible cell cycle exit. C2C12 myoblasts were transfected with control, BRG1, BAF53a or BAF47 siRNAs and cells were labeled with BrdU either after 96 h in DM or 12 h after the switch back to GM (96+12 h GM). We show one representative of 3 independent experiments. Left panel: BrdU indexes ratio of BrdU positive nuclei/DAPI labeled nuclei) are shown in the histogram, error bars represent SEM from 3 quantifications of the presented experiment (*p

    Article Snippet: For ChIP experiments, we used anti-BRG1 (ab110641) and anti-BAF47 (ab12167) from Abcam.

    Techniques: Transfection, Labeling

    SWI/SNF complex subunits are differently involved in skeletal muscle terminal differentiation. A. C2C12 myoblasts were transfected with control, BRG1, BAF53a or BAF47 siRNAs. Immunofluorescence analyses using anti-MCK antibody were performed after 72h in DM. Cells were DAPI-stained prior to fluorescent microscopy analyses (magnification x20). B and C. C2C12 myoblasts were transfected with control, BRG1, BAF53a or BAF47 siRNAs. 24 h post-transfection (0) cells were placed in DM and harvested after 24 h, 48 h or 72 h. B. Total protein extracts were analyzed by WB with the indicated antibodies. *: non-specific bands. For each time point, % of protein siRNA-induced downregulation compared to the same time from control scrambled siRNA is indicated at the bottom of the western blot (see Figure S2 for additional WB analyses). C. Quantification of MCK levels from 3 to 8 independent western blot. Error bars represent standard error of the mean (SEM). For each time point, statistics were calculated compared to the same time point from control. Significative p-values (Student T-test, two tailed, unpaired) are indicated * =

    Journal: PLoS ONE

    Article Title: The SWI/SNF Subunit/Tumor Suppressor BAF47/INI1 Is Essential in Cell Cycle Arrest upon Skeletal Muscle Terminal Differentiation

    doi: 10.1371/journal.pone.0108858

    Figure Lengend Snippet: SWI/SNF complex subunits are differently involved in skeletal muscle terminal differentiation. A. C2C12 myoblasts were transfected with control, BRG1, BAF53a or BAF47 siRNAs. Immunofluorescence analyses using anti-MCK antibody were performed after 72h in DM. Cells were DAPI-stained prior to fluorescent microscopy analyses (magnification x20). B and C. C2C12 myoblasts were transfected with control, BRG1, BAF53a or BAF47 siRNAs. 24 h post-transfection (0) cells were placed in DM and harvested after 24 h, 48 h or 72 h. B. Total protein extracts were analyzed by WB with the indicated antibodies. *: non-specific bands. For each time point, % of protein siRNA-induced downregulation compared to the same time from control scrambled siRNA is indicated at the bottom of the western blot (see Figure S2 for additional WB analyses). C. Quantification of MCK levels from 3 to 8 independent western blot. Error bars represent standard error of the mean (SEM). For each time point, statistics were calculated compared to the same time point from control. Significative p-values (Student T-test, two tailed, unpaired) are indicated * =

    Article Snippet: For ChIP experiments, we used anti-BRG1 (ab110641) and anti-BAF47 (ab12167) from Abcam.

    Techniques: Transfection, Immunofluorescence, Staining, Microscopy, Western Blot, Two Tailed Test

    Downregulation of BAF47 alters muscle terminal differentiation and cell cycle exit. A. BAF47 and BRG1 occupancy at cyclin D1 promoter. ChIP-qPCR analyses of BAF47 and BRG1 in myoblast in proliferating C2C12 cells and at 24 h of differentiation. The immunoprecipitated material was quantified by qPCR, and results are expressed as fold enrichment of the % of Input of BAF47 or BRG1 ChIP over % of Input of the IgG average. Data are represented as mean ±SEM, n = 3. B. Scheme of the timing for samples collection. C2C12 myoblasts were transfected with control, BRG1, BAF53a or BAF47 siRNAs and samples prepared either 24 h after transfection (0) or after 72 h in DM (72) or 10 h after the switch back to GM (72+10 h GM). C. Total protein extracts were analyzed by WB with the indicated antibodies. D. Quantification of cyD1 levels from 3 to 8 independent WB. Data are expressed compared to control 0 h. Error bars represent SEM. For each time point, statistics were calculated compare to the same time point from control. Only significative p-values (Student T-test, two tailed, unpaired) are indicated ** =

    Journal: PLoS ONE

    Article Title: The SWI/SNF Subunit/Tumor Suppressor BAF47/INI1 Is Essential in Cell Cycle Arrest upon Skeletal Muscle Terminal Differentiation

    doi: 10.1371/journal.pone.0108858

    Figure Lengend Snippet: Downregulation of BAF47 alters muscle terminal differentiation and cell cycle exit. A. BAF47 and BRG1 occupancy at cyclin D1 promoter. ChIP-qPCR analyses of BAF47 and BRG1 in myoblast in proliferating C2C12 cells and at 24 h of differentiation. The immunoprecipitated material was quantified by qPCR, and results are expressed as fold enrichment of the % of Input of BAF47 or BRG1 ChIP over % of Input of the IgG average. Data are represented as mean ±SEM, n = 3. B. Scheme of the timing for samples collection. C2C12 myoblasts were transfected with control, BRG1, BAF53a or BAF47 siRNAs and samples prepared either 24 h after transfection (0) or after 72 h in DM (72) or 10 h after the switch back to GM (72+10 h GM). C. Total protein extracts were analyzed by WB with the indicated antibodies. D. Quantification of cyD1 levels from 3 to 8 independent WB. Data are expressed compared to control 0 h. Error bars represent SEM. For each time point, statistics were calculated compare to the same time point from control. Only significative p-values (Student T-test, two tailed, unpaired) are indicated ** =

    Article Snippet: For ChIP experiments, we used anti-BRG1 (ab110641) and anti-BAF47 (ab12167) from Abcam.

    Techniques: Chromatin Immunoprecipitation, Real-time Polymerase Chain Reaction, Immunoprecipitation, Transfection, Western Blot, Two Tailed Test

    BRG1 and BAF47 interact with SWI/SNF and N-CoR-1 complex components in a differentiation-dependent manner. A. Nuclear extracts from proliferating (GM) or differentiating C2C12 myoblasts (24 h DM) were used for immunoprecipitation (IP) with antibodies against BRG1 or BAF47, or with normal rabbit IgG as a negative control. The resulting precipitates were analyzed by WB with the indicated antibodies. Nuclear extracts (Inputs, 1% of input extracts) were loaded to assess endogenous protein levels. *: non-specific IgG band. B. Nuclear extracts from proliferating (GM) or differentiating C2C12 myoblasts (24 h DM) were fractionated on glycerol gradient ranging from 11% (fraction 1) to 33% (fraction 11); (-) empty lane. Fractions were collected and analyzed by WB using the indicated antibodies. Nuclear extracts (Inputs, 2.5% of input extracts) were loaded to assess endogenous protein levels.

    Journal: PLoS ONE

    Article Title: The SWI/SNF Subunit/Tumor Suppressor BAF47/INI1 Is Essential in Cell Cycle Arrest upon Skeletal Muscle Terminal Differentiation

    doi: 10.1371/journal.pone.0108858

    Figure Lengend Snippet: BRG1 and BAF47 interact with SWI/SNF and N-CoR-1 complex components in a differentiation-dependent manner. A. Nuclear extracts from proliferating (GM) or differentiating C2C12 myoblasts (24 h DM) were used for immunoprecipitation (IP) with antibodies against BRG1 or BAF47, or with normal rabbit IgG as a negative control. The resulting precipitates were analyzed by WB with the indicated antibodies. Nuclear extracts (Inputs, 1% of input extracts) were loaded to assess endogenous protein levels. *: non-specific IgG band. B. Nuclear extracts from proliferating (GM) or differentiating C2C12 myoblasts (24 h DM) were fractionated on glycerol gradient ranging from 11% (fraction 1) to 33% (fraction 11); (-) empty lane. Fractions were collected and analyzed by WB using the indicated antibodies. Nuclear extracts (Inputs, 2.5% of input extracts) were loaded to assess endogenous protein levels.

    Article Snippet: For ChIP experiments, we used anti-BRG1 (ab110641) and anti-BAF47 (ab12167) from Abcam.

    Techniques: Immunoprecipitation, Negative Control, Western Blot