Journal: iScience

Article Title: An AIB1 isoform rewires glucocorticoid receptor signaling to promote TNBC progression

doi: 10.1016/j.isci.2026.114768

Figure Lengend Snippet: Mouse AIB1Δ5 is a homolog of human AIB1Δ4 (A) Representative Sashimi plot of RNA sequencing reads skipping exon 5 (blue arrow) in mouse E0771 cells. (B) Expression level of the GR-regulated gene Fkbp5 in E0771 empty vector (E0771-EV) and E0771-AIB1Δ5 overexpressing (E0771-Δ5) cells treated with either 10 nM dex or 1 μM rela for 48 h ( n = 3 technical replicates per condition). Data are represented as mean ± SD. ∗ p < 0.05. ∗∗∗ p < 0.001. p value was calculated using two-tailed unpaired Student’s t-tests. (C) Relative level of AIB1Δ5 mRNA in indicated E0771 samples, normalized to full-length AIB1. Normal lung samples are from control female C57BL/6 mice without E0771 tumor cell injection. Data represent n = 4 biological replicates for E0771 cells grown in culture, n = 3 biological replicates for tumor samples, n = 3 biological replicates for lung metastasis, and n = 3 biological replicates for tumor naive lung tissue. Data are represented as mean ± SD. ∗ p < 0.05. p value was calculated using two-tailed unpaired Student’s t test. (D) Expression level of AIB1 and AIB1Δ5 mRNA using RT-qPCR in the indicated samples. Tumor and immune cells were dissociated from primary tumor and spleen tissues and FACS sorted for CD45 − , CD45 + , or CD45+/CD3+ cells prior to RNA extraction and RT-qPCR. Median plotted as a solid line. Data represent n = 4 biological replicates for E0771 cells grown in culture, n = 8 tumor samples ( n = 4 mice), and n = 3 mouse spleens. ∗∗ p < 0.01. ∗∗∗ p < 0.001. ∗∗∗∗ p < 0.0001. p value was calculated using two-tailed unpaired Student’s t test.

Article Snippet: 24 hours later, whole cell lysate was prepared with NP40 lysis buffer and 1 mg of protein was used for immunoprecipitation with AIB1 antibody (Cell Signaling Technology, 2126).

Techniques: RNA Sequencing, Expressing, Plasmid Preparation, Two Tailed Test, Control, Injection, Quantitative RT-PCR, RNA Extraction

Journal: iScience

Article Title: An AIB1 isoform rewires glucocorticoid receptor signaling to promote TNBC progression

doi: 10.1016/j.isci.2026.114768

Figure Lengend Snippet: Glucocorticoid transcriptional responses are altered by the switch to AIBΔ4 expression in TNBC cells (A) Luciferase assay using MMTV promoter-driven firefly luciferase. HEK 293T cells were transfected with an MMTV promoter-driven firefly construct, a human GR expression vector, and either a pcDNA3 empty vector, pcDNA3-AIB1, or pcDNA3-AIB1Δ4 expression vector. Cells were treated with either vehicle control or 10 nM dex. Luciferase activity was measured 24 h after transfection. Dot blot shows AIB1 protein levels in the three samples treated with dex. Below the dot blot are the protein expression levels normalized to pcDNA3-empty vector. Data are represented as mean ± SD ( n = 3). RLU = relative light unit. ∗∗∗∗ p < 0.0001. p value was calculated using two-tailed unpaired Student’s t-tests. (B) Western blot for GR in DCIS and DCISΔ4 cells after overnight treatment with vehicle, 5 nM dex, or 5 nM dex plus 500 nM RU486. (C) Differentially expressed genes (RNA-seq) of dex (5 nM) treated versus dex (5 nM) plus RU486 (500 nM) treated DCIS or DCISΔ4 cells. All differentially expressed genes had an FDR ≤0.05. Data represent n = 3 biological replicates for each cell line and treatment condition. (D) Bar graph depicting differentially expressed genes in B that are either commonly or uniquely activated or repressed in the DCIS and DCISΔ4 cells. (E) Western blot for GR in DCIS and DCISΔ4 cells after 1 h treatment with vehicle, 5 nM dex or 5 nM dex plus 500 nM RU486. Cells were cultured in media without serum and hydrocortisone for 4 h prior to drug treatment in full serum. (F) Peak overlap from a CUT&RUN experiment with a GR antibody in DCIS and DCISΔ4 cells. Cells were treated for 1 h with dex prior to conducting the CUT&RUN experiment. Data represent n = 3 biological replicates for each cell line. Peaks were called with SEACR in stringent mode and MACS3 with a q-value cutoff of ≤0.05. Consensus peaks were those called in at least 2 of the 3 replicates. (G and H) Categorization of significantly enriched motifs from the GR CUT&RUN experiment in DCIS (E) and DCISΔ4 (F) cells. bZIP = basic leucine zipper; bHLH = basic-helix-loop-helix; HMG = high-mobility group; Zf = zinc finger; NR = nuclear receptor. (I) Heatmap representation of ATAC-seq peaks found in DCIS and DCISΔ4 cells. Cells were treated for 1 h with dex prior to the ATAC-seq experiment. Overlap peaks were called in both cell lines ( n = 87,356). DCIS enriched peaks were only called in the DCIS parental cell line ( n = 5,459), and DCISΔ4 enriched peaks were only called in the DCISΔ4 cell line ( n = 60,715). Data represent n = 3 biological replicates for each cell line. Peaks were called with MACS3 with a q-value cutoff of ≤0.01. Consensus peaks were those called in at least 2 of the 3 replicates. (J) GR CUT&RUN reads (RPKM) in DCIS and DCISΔ4 cells plotted at ATAC-seq peak loci from G.

Article Snippet: 24 hours later, whole cell lysate was prepared with NP40 lysis buffer and 1 mg of protein was used for immunoprecipitation with AIB1 antibody (Cell Signaling Technology, 2126).

Techniques: Expressing, Luciferase, Transfection, Construct, Plasmid Preparation, Control, Activity Assay, Dot Blot, Two Tailed Test, Western Blot, RNA Sequencing, Cell Culture

Journal: iScience

Article Title: An AIB1 isoform rewires glucocorticoid receptor signaling to promote TNBC progression

doi: 10.1016/j.isci.2026.114768

Figure Lengend Snippet: AIB1Δ4 chromatin engagement is affected by cell-cell crosstalk (A) Heatmap representation of peaks called from a CUT&RUN experiment with an AIB1 antibody in DCIS and DCISΔ4 cells. Cells were treated for 1 h with dex prior to conducting the CUT&RUN experiment. Data represent n = 3 biological replicates for each cell line. Peaks were called with SEACR in relaxed mode and MACS3 with a p value cutoff of ≤0.005. Consensus peaks were those called in at least 2 of the 3 replicates. (B) Top ten significantly enriched motifs found in the DCISΔ4 cell line from AIB1 CUT&RUN. (C and D) Peak overlap from a CUT&RUN experiment with an AIB1 antibody in DCISΔ4 and DCISΔ4 unmixed cells (C) and DCIS and DCIS unmixed cells (D). Cells were treated for 1 h with dex prior to conducting the CUT&RUN experiment. Data represent n = 3 biological replicates for each cell line. Peaks were called with SEACR in relaxed mode and MACS3 with a p value cutoff of ≤0.005. Consensus peaks were those called in at least 2 of the 3 replicates. (E) Top six significantly enriched motifs found in the DCISΔ4 unmixed cell line from AIB1 CUT&RUN. (F) Top human Enrichr ChEA 2022 terms for the 467 DCISΔ4 unmixed upregulated genes with dex treatment that also have AIB1 binding in DCISΔ4 unmixed cells (from H). The ChEA dataset is from target genes of transcription factors from published ChIP-chip, ChIP-seq, and other transcription factor binding site profiling studies.

Article Snippet: 24 hours later, whole cell lysate was prepared with NP40 lysis buffer and 1 mg of protein was used for immunoprecipitation with AIB1 antibody (Cell Signaling Technology, 2126).

Techniques: Binding Assay, ChIP-chip, ChIP-sequencing

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Transcriptional Regulation of De Novo Lipogenesis by SIX1 in Liver Cancer Cells.

doi: 10.1002/advs.202404229

Figure Lengend Snippet: Figure 3. SIX1 promotes lipogenic gene expression through association with AIB1 and HBO1. A, B) qRT-PCR (A) and immunoblot (B) analysis of AIB1/HBO1 WT or KO HepG2 cells transfected with SIX1, HBO1, AIB1, or EV as indicated. C) ChIP analysis of SIX1, AIB1, and HBO1 occupancy on the promoters of lipogenic genes in HepG2 cells. Promoter regions of each gene represent the region containing the first or second SIX1 binding site shown in Figure 2B within the gene promoters analyzed. D) Re-ChIP analysis of the occupancy of SIX1 and AIB1 or HBO1 on the indicated lipogenic gene promoters in HepG2 cells. E) ChIP analysis of SIX1, HBO1, AIB1, and histone H3 or H4 acetylation (ac) occupancy on the indicated promoters of lipogenic genes in SIX1, AIB1, or HBO1 KO HepG2 cells. Data shown are mean ± SD of triplicate measurements. Experiments have been repeated 3 times with similar results. A two-sided Student’s t-test was used to compare the means of 2 groups. When more than 2 groups were compared, one-way ANOVA was performed. **p < 0.01 versus respective WT HepG2 cells transfected with empty vector (A). *p < 0.05, **p < 0.01 versus respective normal IgG (C-E).

Article Snippet: The membranes were blocked with 5% skimmed milk for 1 h at room temperature, followed by incubation with primary antibodies including anti-ACLY (Proteintech; Cat# 67166-1-Ig), anti-ACC1 antibody (Proteintech; Cat# 21923-1-AP), anti-FASN antibody (Proteintech; Cat# 10624-2-AP), anti-SCD1 antibody (Proteintech; Cat# 28678-1-AP), anti-SIX1 antibody (Proteintech; Cat# 10709-1-AP), anti-AIB1 antibody (Santa Cruz Biotechnology; Cat# sc-9119), anti-HBO1 antibody (Proteintech; Cat# 13751-1-AP), and anti-β-actin (Santa Cruz Biotechnology; Cat# sc-47778HRP) at room temperature for 2 h or overnight at 4 °C.

Techniques: Gene Expression, Quantitative RT-PCR, Western Blot, Transfection, Binding Assay, Plasmid Preparation

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Transcriptional Regulation of De Novo Lipogenesis by SIX1 in Liver Cancer Cells.

doi: 10.1002/advs.202404229

Figure Lengend Snippet: Figure 4. SIX1 is an insulin-responsive gene and stimulates de novo lipogenesis. A) Lipid droplets (LDs) were visualized using BODIPY 493/503 in SIX1 WT or KO HepG2 cells or SIX1 KO HepG2 cells transfected with SIX1. Scale bar, 20 μm. B) Long-chain fatty acid levels were analyzed by liquid chromatography-mass spectrometry in cells from (A). C) Triglyceride and total cholesterol levels were measured in cells from (A). D-F) Lipid droplets (D), long-chain fatty acid levels (E), and triglyceride and total cholesterol levels (F) were analyzed in WT or HBO1/AIB1/SCD1 KO HepG2 cells transfected with SIX1 or EV. G) Immunoblot analysis of SIX1 WT or KO HepG2 cells treated with insulin (100 nM) or linsitinib (1.0 μM). H, I) Long-chain fatty

Article Snippet: The membranes were blocked with 5% skimmed milk for 1 h at room temperature, followed by incubation with primary antibodies including anti-ACLY (Proteintech; Cat# 67166-1-Ig), anti-ACC1 antibody (Proteintech; Cat# 21923-1-AP), anti-FASN antibody (Proteintech; Cat# 10624-2-AP), anti-SCD1 antibody (Proteintech; Cat# 28678-1-AP), anti-SIX1 antibody (Proteintech; Cat# 10709-1-AP), anti-AIB1 antibody (Santa Cruz Biotechnology; Cat# sc-9119), anti-HBO1 antibody (Proteintech; Cat# 13751-1-AP), and anti-β-actin (Santa Cruz Biotechnology; Cat# sc-47778HRP) at room temperature for 2 h or overnight at 4 °C.

Techniques: Transfection, Liquid Chromatography, Mass Spectrometry, Western Blot

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Transcriptional Regulation of De Novo Lipogenesis by SIX1 in Liver Cancer Cells.

doi: 10.1002/advs.202404229

Figure Lengend Snippet: Figure 7. Clinical relevance of the DGUOK-AS1/miR-145-5p/SIX1 axis in liver cancer. A) Representative IHC of 73 liver cancer patients. SIX1 and SCD1 were assessed by IHC, and DGUOK-AS1 and miR-145-5p by FISH. Scale bar: 100 μm. The correlations among DGUOK-AS1, miR-145-5p, SCD1, and SIX1 were analyzed as indicated. Case 1 and case 2 refer to 2 representative samples categorized by low and high expression of SIX1. Data was analyzed by Spearman’s Rank Correlation test. B) DGUOK-AS1 expression in 73 cancerous liver tissues and matched adjacent normal liver tissues was determined by FISH. The DGUOK-AS1 expression levels were plotted and compared between normal and cancer tissues (Mann-Whitney U test). C) The disease-free and overall survival curves related to low and high expression of DGUOK-AS1 were analyzed in 73 liver cancer patients from (A) using the Kaplan-Meier method. D) A proposed model underlying the role of the insulin/DGUOK-AS1/miR-145-5p/SIX1 axis in de novo lipogenesis and liver tumor growth and metastasis. Insulin stimulates the expression of DGUOK-AS1, thus sponging miR-145-5p. Inhibition of miR-145-5p promotes expression of SIX1, which recruits histone acetyltransferases AIB1 and HBO1 to induce expression of lipogenic genes (ACLY, FASN, and SCD1). Induction of lipogenic gene expression promotes hepatic lipogenesis and liver tumor growth and metastasis. FAs, fatty acids. TGs, triglycerides. LDs, lipid droplets.

Article Snippet: The membranes were blocked with 5% skimmed milk for 1 h at room temperature, followed by incubation with primary antibodies including anti-ACLY (Proteintech; Cat# 67166-1-Ig), anti-ACC1 antibody (Proteintech; Cat# 21923-1-AP), anti-FASN antibody (Proteintech; Cat# 10624-2-AP), anti-SCD1 antibody (Proteintech; Cat# 28678-1-AP), anti-SIX1 antibody (Proteintech; Cat# 10709-1-AP), anti-AIB1 antibody (Santa Cruz Biotechnology; Cat# sc-9119), anti-HBO1 antibody (Proteintech; Cat# 13751-1-AP), and anti-β-actin (Santa Cruz Biotechnology; Cat# sc-47778HRP) at room temperature for 2 h or overnight at 4 °C.

Techniques: Expressing, MANN-WHITNEY, Inhibition, Gene Expression

Journal: PLoS ONE

Article Title: Proteomic Analysis of Pathways Involved in Estrogen-Induced Growth and Apoptosis of Breast Cancer Cells

doi: 10.1371/journal.pone.0020410

Figure Lengend Snippet: ( A ) The experimental paradigm. The differential responses to estradiol (E2) treatment of MCF-7 (cell growth) and long-term estrogen deprived MCF-7:5C cells (apoptosis) are indicated. Proteomics profiles of the two cell lines at baseline and after a brief (2 h) E2 treatment were generated using immunoprecipitations (IP). Proteins interacting with AIB1 or phosphotyrosine containing protein complexes were isolated by IP followed by mass spectrometry. Data were then subjected to an integrated bioinformatics analysis of signaling pathways and protein networks. ( B,C ) Reversal of E2-dependent effects on MCF-7 and MCF-7:5C after depletion of endogenous AIB1 protein using two different lentiviral shRNAs. MCF-7 or MCF-7:5C cells were infected with lentiviral particles expressing control or AIB1-targeting shRNAs. (B) RNAi-mediated knockdown was assayed by Western blot analysis for AIB1 relative to an actin loading control. (C) Cell growth was assayed 6 days after plating without or with E2. The E2 effect is shown relative to the respective untreated controls (mean ±S.E.M.). Closed circles: control shRNA; Open circles (red): AIB1 shRNA. #, p<0.05 E2 treatment effect vs. no treatment in control shRNA cells; *, p<0.05 E2 treatment effect in control shRNA cells vs. E2 treatment in AIB1 depleted cells. Representative data from one of at least three independent experiments are shown.

Article Snippet: Western blot analyses were done as previously described , using a monoclonal antibody for AIB1 (SRC3; clone 5E11, Cell Signaling).

Techniques: Generated, Isolation, Mass Spectrometry, Infection, Expressing, Western Blot, shRNA

Journal: PLoS ONE

Article Title: Proteomic Analysis of Pathways Involved in Estrogen-Induced Growth and Apoptosis of Breast Cancer Cells

doi: 10.1371/journal.pone.0020410

Figure Lengend Snippet: Venn diagrams of proteins identified from anti-AIB1 ( A,C ) or anti-pY IP ( B,D ) experimental groups. ( C,D ) Proteins in combined AIB1-IP or pY-IP data sets. Individual proteins and subgroups are shown in & .

Article Snippet: Western blot analyses were done as previously described , using a monoclonal antibody for AIB1 (SRC3; clone 5E11, Cell Signaling).

Techniques:

Journal: PLoS ONE

Article Title: Proteomic Analysis of Pathways Involved in Estrogen-Induced Growth and Apoptosis of Breast Cancer Cells

doi: 10.1371/journal.pone.0020410

Figure Lengend Snippet: The thick grey line in the middle provides an arbitrary boundary between the pathways. Anti-AIB1 immunoprecipitated (AIB1-IPed) and anti-pY-immunoprecipitated proteins (pY-IPed) are indicated by red or green circles respectively (keys at the bottom). The blue circled proteins are AIB1-IPed proteins from MCF-7 (CALM1) or MCF-7:5C cells (β-catenin) under both E2-treated and untreated conditions; the purple circled one (ITPR3) is an AIB1-IPed protein from both cells only under E2 treated condition, while the yellow circled one (TYK2) is an AIB1-IPed protein from both cells under both E2 treated and untreated conditions. Proteins circled in grey are from known canonical pathways (e.g. ERK in cell growth or BAD in apoptosis) but not identified here. Solid line arrows indicate direct interactions (e.g. CDK1 phosphorylates Rap1GAP) or translocations (e.g. catenins) of proteins, while dashed arrows indicate indirect actions of proteins (e.g. AKT activate MEK through several steps). Hammer-ended lines indicate inhibitory effects on the target. Detailled pathways are shown in , , , .

Article Snippet: Western blot analyses were done as previously described , using a monoclonal antibody for AIB1 (SRC3; clone 5E11, Cell Signaling).

Techniques: Immunoprecipitation

Journal: PLoS ONE

Article Title: Proteomic Analysis of Pathways Involved in Estrogen-Induced Growth and Apoptosis of Breast Cancer Cells

doi: 10.1371/journal.pone.0020410

Figure Lengend Snippet: ( A ) The experimental paradigm. The differential responses to estradiol (E2) treatment of MCF-7 (cell growth) and long-term estrogen deprived MCF-7:5C cells (apoptosis) are indicated. Proteomics profiles of the two cell lines at baseline and after a brief (2 h) E2 treatment were generated using immunoprecipitations (IP). Proteins interacting with AIB1 or phosphotyrosine containing protein complexes were isolated by IP followed by mass spectrometry. Data were then subjected to an integrated bioinformatics analysis of signaling pathways and protein networks. ( B,C ) Reversal of E2-dependent effects on MCF-7 and MCF-7:5C after depletion of endogenous AIB1 protein using two different lentiviral shRNAs. MCF-7 or MCF-7:5C cells were infected with lentiviral particles expressing control or AIB1-targeting shRNAs. (B) RNAi-mediated knockdown was assayed by Western blot analysis for AIB1 relative to an actin loading control. (C) Cell growth was assayed 6 days after plating without or with E2. The E2 effect is shown relative to the respective untreated controls (mean ±S.E.M.). Closed circles: control shRNA; Open circles (red): AIB1 shRNA. #, p<0.05 E2 treatment effect vs. no treatment in control shRNA cells; *, p<0.05 E2 treatment effect in control shRNA cells vs. E2 treatment in AIB1 depleted cells. Representative data from one of at least three independent experiments are shown.

Article Snippet: For the mass spectrometry analysis, protein lysates from cells treated for 2 hours with E2 or vehicle were subjected to immunoprecipitation using gamma-bind G-Sepharose beads and an anti-AIB1 monoclonal antibody (BD Biosciences) as described or an anti-phosphotyrosine monoclonal antibody (4G-10, Millipore).

Techniques: Generated, Isolation, Mass Spectrometry, Infection, Expressing, Western Blot, shRNA

Journal: PLoS ONE

Article Title: Proteomic Analysis of Pathways Involved in Estrogen-Induced Growth and Apoptosis of Breast Cancer Cells

doi: 10.1371/journal.pone.0020410

Figure Lengend Snippet: Venn diagrams of proteins identified from anti-AIB1 ( A,C ) or anti-pY IP ( B,D ) experimental groups. ( C,D ) Proteins in combined AIB1-IP or pY-IP data sets. Individual proteins and subgroups are shown in & .

Article Snippet: For the mass spectrometry analysis, protein lysates from cells treated for 2 hours with E2 or vehicle were subjected to immunoprecipitation using gamma-bind G-Sepharose beads and an anti-AIB1 monoclonal antibody (BD Biosciences) as described or an anti-phosphotyrosine monoclonal antibody (4G-10, Millipore).

Techniques:

Journal: PLoS ONE

Article Title: Proteomic Analysis of Pathways Involved in Estrogen-Induced Growth and Apoptosis of Breast Cancer Cells

doi: 10.1371/journal.pone.0020410

Figure Lengend Snippet: The thick grey line in the middle provides an arbitrary boundary between the pathways. Anti-AIB1 immunoprecipitated (AIB1-IPed) and anti-pY-immunoprecipitated proteins (pY-IPed) are indicated by red or green circles respectively (keys at the bottom). The blue circled proteins are AIB1-IPed proteins from MCF-7 (CALM1) or MCF-7:5C cells (β-catenin) under both E2-treated and untreated conditions; the purple circled one (ITPR3) is an AIB1-IPed protein from both cells only under E2 treated condition, while the yellow circled one (TYK2) is an AIB1-IPed protein from both cells under both E2 treated and untreated conditions. Proteins circled in grey are from known canonical pathways (e.g. ERK in cell growth or BAD in apoptosis) but not identified here. Solid line arrows indicate direct interactions (e.g. CDK1 phosphorylates Rap1GAP) or translocations (e.g. catenins) of proteins, while dashed arrows indicate indirect actions of proteins (e.g. AKT activate MEK through several steps). Hammer-ended lines indicate inhibitory effects on the target. Detailled pathways are shown in , , , .

Article Snippet: For the mass spectrometry analysis, protein lysates from cells treated for 2 hours with E2 or vehicle were subjected to immunoprecipitation using gamma-bind G-Sepharose beads and an anti-AIB1 monoclonal antibody (BD Biosciences) as described or an anti-phosphotyrosine monoclonal antibody (4G-10, Millipore).

Techniques: Immunoprecipitation

Journal: PLoS ONE

Article Title: Proteomic Analysis of Pathways Involved in Estrogen-Induced Growth and Apoptosis of Breast Cancer Cells

doi: 10.1371/journal.pone.0020410

Figure Lengend Snippet: ( A ) The experimental paradigm. The differential responses to estradiol (E2) treatment of MCF-7 (cell growth) and long-term estrogen deprived MCF-7:5C cells (apoptosis) are indicated. Proteomics profiles of the two cell lines at baseline and after a brief (2 h) E2 treatment were generated using immunoprecipitations (IP). Proteins interacting with AIB1 or phosphotyrosine containing protein complexes were isolated by IP followed by mass spectrometry. Data were then subjected to an integrated bioinformatics analysis of signaling pathways and protein networks. ( B,C ) Reversal of E2-dependent effects on MCF-7 and MCF-7:5C after depletion of endogenous AIB1 protein using two different lentiviral shRNAs. MCF-7 or MCF-7:5C cells were infected with lentiviral particles expressing control or AIB1-targeting shRNAs. (B) RNAi-mediated knockdown was assayed by Western blot analysis for AIB1 relative to an actin loading control. (C) Cell growth was assayed 6 days after plating without or with E2. The E2 effect is shown relative to the respective untreated controls (mean ±S.E.M.). Closed circles: control shRNA; Open circles (red): AIB1 shRNA. #, p<0.05 E2 treatment effect vs. no treatment in control shRNA cells; *, p<0.05 E2 treatment effect in control shRNA cells vs. E2 treatment in AIB1 depleted cells. Representative data from one of at least three independent experiments are shown.

Article Snippet: The AIB1(1) shRNA was derived from an siRNA for AIB1 previously described , and the AIB1(2) shRNA was from Sigma (TRCN0000019703).

Techniques: Generated, Isolation, Mass Spectrometry, Infection, Expressing, Western Blot, shRNA

Journal: PLoS ONE

Article Title: Proteomic Analysis of Pathways Involved in Estrogen-Induced Growth and Apoptosis of Breast Cancer Cells

doi: 10.1371/journal.pone.0020410

Figure Lengend Snippet: Venn diagrams of proteins identified from anti-AIB1 ( A,C ) or anti-pY IP ( B,D ) experimental groups. ( C,D ) Proteins in combined AIB1-IP or pY-IP data sets. Individual proteins and subgroups are shown in & .

Article Snippet: The AIB1(1) shRNA was derived from an siRNA for AIB1 previously described , and the AIB1(2) shRNA was from Sigma (TRCN0000019703).

Techniques:

Journal: PLoS ONE

Article Title: Proteomic Analysis of Pathways Involved in Estrogen-Induced Growth and Apoptosis of Breast Cancer Cells

doi: 10.1371/journal.pone.0020410

Figure Lengend Snippet: The thick grey line in the middle provides an arbitrary boundary between the pathways. Anti-AIB1 immunoprecipitated (AIB1-IPed) and anti-pY-immunoprecipitated proteins (pY-IPed) are indicated by red or green circles respectively (keys at the bottom). The blue circled proteins are AIB1-IPed proteins from MCF-7 (CALM1) or MCF-7:5C cells (β-catenin) under both E2-treated and untreated conditions; the purple circled one (ITPR3) is an AIB1-IPed protein from both cells only under E2 treated condition, while the yellow circled one (TYK2) is an AIB1-IPed protein from both cells under both E2 treated and untreated conditions. Proteins circled in grey are from known canonical pathways (e.g. ERK in cell growth or BAD in apoptosis) but not identified here. Solid line arrows indicate direct interactions (e.g. CDK1 phosphorylates Rap1GAP) or translocations (e.g. catenins) of proteins, while dashed arrows indicate indirect actions of proteins (e.g. AKT activate MEK through several steps). Hammer-ended lines indicate inhibitory effects on the target. Detailled pathways are shown in , , , .

Article Snippet: The AIB1(1) shRNA was derived from an siRNA for AIB1 previously described , and the AIB1(2) shRNA was from Sigma (TRCN0000019703).

Techniques: Immunoprecipitation