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AF6 regulates the expression of MHC II by modulating the expression of <t>STAT1</t> in intestinal epithelial cells (IECs) (A) The protein levels of components of the IFN-γ-related signaling pathway were detected by immunoblotting of colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (B) qPCR was used to assess STAT1 transcript levels in colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (C and D) (C) The mRNA levels of STAT1 and its downstream target genes (as assessed by qPCR) and (D) the protein levels of STAT1 and proteins encoded by its downstream target genes (as assessed by immunoblotting) were determined in organoids generated from colon tissues of Af6 f/f and Af6 ΔIEC mice, as measured before and after IFN-γ treatment. (E) Co-immunoprecipitation (co-IP) was used to detect endogenous interactions between AF6 and IRF1 in IECs. (F) 293T cells were co-transfected with constructs encoding hemagglutinin-tagged AF6 (HA-AF6) and Flag peptide-tagged IRF1 (Flag-IRF1); their interaction domains were detected by co-IP. (G) Immunoblotting was used to assess the expression of IRF1 and proteins encoded by its downstream genes in HT29 cells with or without IFN-γ exposure. (H) HT29 cells were transfected with constructs encoding AF6 with no or 3 nuclear localization signaling (NLS) domains (ΔNLS-AF6 and 3×NLS-AF6, respectively), and the expression and localization of IRF1 and proteins encoded by its downstream genes were assessed by immunoblotting following the separation of the nuclear and cytoplasmic fractions. Data are expressed as mean ± SEM. Pairwise comparisons between groups were conducted using two-tailed non-paired Student’s t tests. p -values were shown in the panel, and p < 0.05 indicates a significant difference.
Phospho Stat1 (Tyr701) (58d6) Rabbit Mab, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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AF6 regulates the expression of MHC II by modulating the expression of <t>STAT1</t> in intestinal epithelial cells (IECs) (A) The protein levels of components of the IFN-γ-related signaling pathway were detected by immunoblotting of colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (B) qPCR was used to assess STAT1 transcript levels in colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (C and D) (C) The mRNA levels of STAT1 and its downstream target genes (as assessed by qPCR) and (D) the protein levels of STAT1 and proteins encoded by its downstream target genes (as assessed by immunoblotting) were determined in organoids generated from colon tissues of Af6 f/f and Af6 ΔIEC mice, as measured before and after IFN-γ treatment. (E) Co-immunoprecipitation (co-IP) was used to detect endogenous interactions between AF6 and IRF1 in IECs. (F) 293T cells were co-transfected with constructs encoding hemagglutinin-tagged AF6 (HA-AF6) and Flag peptide-tagged IRF1 (Flag-IRF1); their interaction domains were detected by co-IP. (G) Immunoblotting was used to assess the expression of IRF1 and proteins encoded by its downstream genes in HT29 cells with or without IFN-γ exposure. (H) HT29 cells were transfected with constructs encoding AF6 with no or 3 nuclear localization signaling (NLS) domains (ΔNLS-AF6 and 3×NLS-AF6, respectively), and the expression and localization of IRF1 and proteins encoded by its downstream genes were assessed by immunoblotting following the separation of the nuclear and cytoplasmic fractions. Data are expressed as mean ± SEM. Pairwise comparisons between groups were conducted using two-tailed non-paired Student’s t tests. p -values were shown in the panel, and p < 0.05 indicates a significant difference.
P Stat1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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phospho stat1 tyr701  (Cell Signaling Technology Inc)
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AF6 regulates the expression of MHC II by modulating the expression of <t>STAT1</t> in intestinal epithelial cells (IECs) (A) The protein levels of components of the IFN-γ-related signaling pathway were detected by immunoblotting of colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (B) qPCR was used to assess STAT1 transcript levels in colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (C and D) (C) The mRNA levels of STAT1 and its downstream target genes (as assessed by qPCR) and (D) the protein levels of STAT1 and proteins encoded by its downstream target genes (as assessed by immunoblotting) were determined in organoids generated from colon tissues of Af6 f/f and Af6 ΔIEC mice, as measured before and after IFN-γ treatment. (E) Co-immunoprecipitation (co-IP) was used to detect endogenous interactions between AF6 and IRF1 in IECs. (F) 293T cells were co-transfected with constructs encoding hemagglutinin-tagged AF6 (HA-AF6) and Flag peptide-tagged IRF1 (Flag-IRF1); their interaction domains were detected by co-IP. (G) Immunoblotting was used to assess the expression of IRF1 and proteins encoded by its downstream genes in HT29 cells with or without IFN-γ exposure. (H) HT29 cells were transfected with constructs encoding AF6 with no or 3 nuclear localization signaling (NLS) domains (ΔNLS-AF6 and 3×NLS-AF6, respectively), and the expression and localization of IRF1 and proteins encoded by its downstream genes were assessed by immunoblotting following the separation of the nuclear and cytoplasmic fractions. Data are expressed as mean ± SEM. Pairwise comparisons between groups were conducted using two-tailed non-paired Student’s t tests. p -values were shown in the panel, and p < 0.05 indicates a significant difference.
Phospho Stat1 Tyr701, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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AF6 regulates the expression of MHC II by modulating the expression of <t>STAT1</t> in intestinal epithelial cells (IECs) (A) The protein levels of components of the IFN-γ-related signaling pathway were detected by immunoblotting of colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (B) qPCR was used to assess STAT1 transcript levels in colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (C and D) (C) The mRNA levels of STAT1 and its downstream target genes (as assessed by qPCR) and (D) the protein levels of STAT1 and proteins encoded by its downstream target genes (as assessed by immunoblotting) were determined in organoids generated from colon tissues of Af6 f/f and Af6 ΔIEC mice, as measured before and after IFN-γ treatment. (E) Co-immunoprecipitation (co-IP) was used to detect endogenous interactions between AF6 and IRF1 in IECs. (F) 293T cells were co-transfected with constructs encoding hemagglutinin-tagged AF6 (HA-AF6) and Flag peptide-tagged IRF1 (Flag-IRF1); their interaction domains were detected by co-IP. (G) Immunoblotting was used to assess the expression of IRF1 and proteins encoded by its downstream genes in HT29 cells with or without IFN-γ exposure. (H) HT29 cells were transfected with constructs encoding AF6 with no or 3 nuclear localization signaling (NLS) domains (ΔNLS-AF6 and 3×NLS-AF6, respectively), and the expression and localization of IRF1 and proteins encoded by its downstream genes were assessed by immunoblotting following the separation of the nuclear and cytoplasmic fractions. Data are expressed as mean ± SEM. Pairwise comparisons between groups were conducted using two-tailed non-paired Student’s t tests. p -values were shown in the panel, and p < 0.05 indicates a significant difference.
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stat2 d9j7l cell signaling technology ab 2799824 cat  (Cell Signaling Technology Inc)
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AF6 regulates the expression of MHC II by modulating the expression of <t>STAT1</t> in intestinal epithelial cells (IECs) (A) The protein levels of components of the IFN-γ-related signaling pathway were detected by immunoblotting of colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (B) qPCR was used to assess STAT1 transcript levels in colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (C and D) (C) The mRNA levels of STAT1 and its downstream target genes (as assessed by qPCR) and (D) the protein levels of STAT1 and proteins encoded by its downstream target genes (as assessed by immunoblotting) were determined in organoids generated from colon tissues of Af6 f/f and Af6 ΔIEC mice, as measured before and after IFN-γ treatment. (E) Co-immunoprecipitation (co-IP) was used to detect endogenous interactions between AF6 and IRF1 in IECs. (F) 293T cells were co-transfected with constructs encoding hemagglutinin-tagged AF6 (HA-AF6) and Flag peptide-tagged IRF1 (Flag-IRF1); their interaction domains were detected by co-IP. (G) Immunoblotting was used to assess the expression of IRF1 and proteins encoded by its downstream genes in HT29 cells with or without IFN-γ exposure. (H) HT29 cells were transfected with constructs encoding AF6 with no or 3 nuclear localization signaling (NLS) domains (ΔNLS-AF6 and 3×NLS-AF6, respectively), and the expression and localization of IRF1 and proteins encoded by its downstream genes were assessed by immunoblotting following the separation of the nuclear and cytoplasmic fractions. Data are expressed as mean ± SEM. Pairwise comparisons between groups were conducted using two-tailed non-paired Student’s t tests. p -values were shown in the panel, and p < 0.05 indicates a significant difference.
Stat2 D9j7l Cell Signaling Technology Ab 2799824 Cat, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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58d6 immunoblot cell signaling technology 9167 stat2 d9j7l  (Cell Signaling Technology Inc)
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AF6 regulates the expression of MHC II by modulating the expression of <t>STAT1</t> in intestinal epithelial cells (IECs) (A) The protein levels of components of the IFN-γ-related signaling pathway were detected by immunoblotting of colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (B) qPCR was used to assess STAT1 transcript levels in colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (C and D) (C) The mRNA levels of STAT1 and its downstream target genes (as assessed by qPCR) and (D) the protein levels of STAT1 and proteins encoded by its downstream target genes (as assessed by immunoblotting) were determined in organoids generated from colon tissues of Af6 f/f and Af6 ΔIEC mice, as measured before and after IFN-γ treatment. (E) Co-immunoprecipitation (co-IP) was used to detect endogenous interactions between AF6 and IRF1 in IECs. (F) 293T cells were co-transfected with constructs encoding hemagglutinin-tagged AF6 (HA-AF6) and Flag peptide-tagged IRF1 (Flag-IRF1); their interaction domains were detected by co-IP. (G) Immunoblotting was used to assess the expression of IRF1 and proteins encoded by its downstream genes in HT29 cells with or without IFN-γ exposure. (H) HT29 cells were transfected with constructs encoding AF6 with no or 3 nuclear localization signaling (NLS) domains (ΔNLS-AF6 and 3×NLS-AF6, respectively), and the expression and localization of IRF1 and proteins encoded by its downstream genes were assessed by immunoblotting following the separation of the nuclear and cytoplasmic fractions. Data are expressed as mean ± SEM. Pairwise comparisons between groups were conducted using two-tailed non-paired Student’s t tests. p -values were shown in the panel, and p < 0.05 indicates a significant difference.
58d6 Immunoblot Cell Signaling Technology 9167 Stat2 D9j7l, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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stat1  (Cell Signaling Technology Inc)
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AF6 regulates the expression of MHC II by modulating the expression of <t>STAT1</t> in intestinal epithelial cells (IECs) (A) The protein levels of components of the IFN-γ-related signaling pathway were detected by immunoblotting of colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (B) qPCR was used to assess STAT1 transcript levels in colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (C and D) (C) The mRNA levels of STAT1 and its downstream target genes (as assessed by qPCR) and (D) the protein levels of STAT1 and proteins encoded by its downstream target genes (as assessed by immunoblotting) were determined in organoids generated from colon tissues of Af6 f/f and Af6 ΔIEC mice, as measured before and after IFN-γ treatment. (E) Co-immunoprecipitation (co-IP) was used to detect endogenous interactions between AF6 and IRF1 in IECs. (F) 293T cells were co-transfected with constructs encoding hemagglutinin-tagged AF6 (HA-AF6) and Flag peptide-tagged IRF1 (Flag-IRF1); their interaction domains were detected by co-IP. (G) Immunoblotting was used to assess the expression of IRF1 and proteins encoded by its downstream genes in HT29 cells with or without IFN-γ exposure. (H) HT29 cells were transfected with constructs encoding AF6 with no or 3 nuclear localization signaling (NLS) domains (ΔNLS-AF6 and 3×NLS-AF6, respectively), and the expression and localization of IRF1 and proteins encoded by its downstream genes were assessed by immunoblotting following the separation of the nuclear and cytoplasmic fractions. Data are expressed as mean ± SEM. Pairwise comparisons between groups were conducted using two-tailed non-paired Student’s t tests. p -values were shown in the panel, and p < 0.05 indicates a significant difference.
Stat1, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 98/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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AF6 regulates the expression of MHC II by modulating the expression of <t>STAT1</t> in intestinal epithelial cells (IECs) (A) The protein levels of components of the IFN-γ-related signaling pathway were detected by immunoblotting of colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (B) qPCR was used to assess STAT1 transcript levels in colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (C and D) (C) The mRNA levels of STAT1 and its downstream target genes (as assessed by qPCR) and (D) the protein levels of STAT1 and proteins encoded by its downstream target genes (as assessed by immunoblotting) were determined in organoids generated from colon tissues of Af6 f/f and Af6 ΔIEC mice, as measured before and after IFN-γ treatment. (E) Co-immunoprecipitation (co-IP) was used to detect endogenous interactions between AF6 and IRF1 in IECs. (F) 293T cells were co-transfected with constructs encoding hemagglutinin-tagged AF6 (HA-AF6) and Flag peptide-tagged IRF1 (Flag-IRF1); their interaction domains were detected by co-IP. (G) Immunoblotting was used to assess the expression of IRF1 and proteins encoded by its downstream genes in HT29 cells with or without IFN-γ exposure. (H) HT29 cells were transfected with constructs encoding AF6 with no or 3 nuclear localization signaling (NLS) domains (ΔNLS-AF6 and 3×NLS-AF6, respectively), and the expression and localization of IRF1 and proteins encoded by its downstream genes were assessed by immunoblotting following the separation of the nuclear and cytoplasmic fractions. Data are expressed as mean ± SEM. Pairwise comparisons between groups were conducted using two-tailed non-paired Student’s t tests. p -values were shown in the panel, and p < 0.05 indicates a significant difference.
7649t, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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58d6 rabbit mab cell signaling technology 9167l p stat1 s727  (Cell Signaling Technology Inc)
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AF6 regulates the expression of MHC II by modulating the expression of <t>STAT1</t> in intestinal epithelial cells (IECs) (A) The protein levels of components of the IFN-γ-related signaling pathway were detected by immunoblotting of colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (B) qPCR was used to assess STAT1 transcript levels in colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (C and D) (C) The mRNA levels of STAT1 and its downstream target genes (as assessed by qPCR) and (D) the protein levels of STAT1 and proteins encoded by its downstream target genes (as assessed by immunoblotting) were determined in organoids generated from colon tissues of Af6 f/f and Af6 ΔIEC mice, as measured before and after IFN-γ treatment. (E) Co-immunoprecipitation (co-IP) was used to detect endogenous interactions between AF6 and IRF1 in IECs. (F) 293T cells were co-transfected with constructs encoding hemagglutinin-tagged AF6 (HA-AF6) and Flag peptide-tagged IRF1 (Flag-IRF1); their interaction domains were detected by co-IP. (G) Immunoblotting was used to assess the expression of IRF1 and proteins encoded by its downstream genes in HT29 cells with or without IFN-γ exposure. (H) HT29 cells were transfected with constructs encoding AF6 with no or 3 nuclear localization signaling (NLS) domains (ΔNLS-AF6 and 3×NLS-AF6, respectively), and the expression and localization of IRF1 and proteins encoded by its downstream genes were assessed by immunoblotting following the separation of the nuclear and cytoplasmic fractions. Data are expressed as mean ± SEM. Pairwise comparisons between groups were conducted using two-tailed non-paired Student’s t tests. p -values were shown in the panel, and p < 0.05 indicates a significant difference.
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7649 stat1 rabbit  (Cell Signaling Technology Inc)
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AF6 regulates the expression of MHC II by modulating the expression of <t>STAT1</t> in intestinal epithelial cells (IECs) (A) The protein levels of components of the IFN-γ-related signaling pathway were detected by immunoblotting of colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (B) qPCR was used to assess STAT1 transcript levels in colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (C and D) (C) The mRNA levels of STAT1 and its downstream target genes (as assessed by qPCR) and (D) the protein levels of STAT1 and proteins encoded by its downstream target genes (as assessed by immunoblotting) were determined in organoids generated from colon tissues of Af6 f/f and Af6 ΔIEC mice, as measured before and after IFN-γ treatment. (E) Co-immunoprecipitation (co-IP) was used to detect endogenous interactions between AF6 and IRF1 in IECs. (F) 293T cells were co-transfected with constructs encoding hemagglutinin-tagged AF6 (HA-AF6) and Flag peptide-tagged IRF1 (Flag-IRF1); their interaction domains were detected by co-IP. (G) Immunoblotting was used to assess the expression of IRF1 and proteins encoded by its downstream genes in HT29 cells with or without IFN-γ exposure. (H) HT29 cells were transfected with constructs encoding AF6 with no or 3 nuclear localization signaling (NLS) domains (ΔNLS-AF6 and 3×NLS-AF6, respectively), and the expression and localization of IRF1 and proteins encoded by its downstream genes were assessed by immunoblotting following the separation of the nuclear and cytoplasmic fractions. Data are expressed as mean ± SEM. Pairwise comparisons between groups were conducted using two-tailed non-paired Student’s t tests. p -values were shown in the panel, and p < 0.05 indicates a significant difference.
7649 Stat1 Rabbit, supplied by Cell Signaling Technology Inc, used in various techniques. Bioz Stars score: 96/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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AF6 regulates the expression of MHC II by modulating the expression of STAT1 in intestinal epithelial cells (IECs) (A) The protein levels of components of the IFN-γ-related signaling pathway were detected by immunoblotting of colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (B) qPCR was used to assess STAT1 transcript levels in colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (C and D) (C) The mRNA levels of STAT1 and its downstream target genes (as assessed by qPCR) and (D) the protein levels of STAT1 and proteins encoded by its downstream target genes (as assessed by immunoblotting) were determined in organoids generated from colon tissues of Af6 f/f and Af6 ΔIEC mice, as measured before and after IFN-γ treatment. (E) Co-immunoprecipitation (co-IP) was used to detect endogenous interactions between AF6 and IRF1 in IECs. (F) 293T cells were co-transfected with constructs encoding hemagglutinin-tagged AF6 (HA-AF6) and Flag peptide-tagged IRF1 (Flag-IRF1); their interaction domains were detected by co-IP. (G) Immunoblotting was used to assess the expression of IRF1 and proteins encoded by its downstream genes in HT29 cells with or without IFN-γ exposure. (H) HT29 cells were transfected with constructs encoding AF6 with no or 3 nuclear localization signaling (NLS) domains (ΔNLS-AF6 and 3×NLS-AF6, respectively), and the expression and localization of IRF1 and proteins encoded by its downstream genes were assessed by immunoblotting following the separation of the nuclear and cytoplasmic fractions. Data are expressed as mean ± SEM. Pairwise comparisons between groups were conducted using two-tailed non-paired Student’s t tests. p -values were shown in the panel, and p < 0.05 indicates a significant difference.

Journal: iScience

Article Title: AF6 regulates intestinal IgA via crosstalk between intestinal epithelial cells and immune cells in inflammatory bowel disease

doi: 10.1016/j.isci.2025.112658

Figure Lengend Snippet: AF6 regulates the expression of MHC II by modulating the expression of STAT1 in intestinal epithelial cells (IECs) (A) The protein levels of components of the IFN-γ-related signaling pathway were detected by immunoblotting of colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (B) qPCR was used to assess STAT1 transcript levels in colon tissues from Af6 f/f and Af6 ΔIEC mice maintained on 2.5% DSS for 7 days. (C and D) (C) The mRNA levels of STAT1 and its downstream target genes (as assessed by qPCR) and (D) the protein levels of STAT1 and proteins encoded by its downstream target genes (as assessed by immunoblotting) were determined in organoids generated from colon tissues of Af6 f/f and Af6 ΔIEC mice, as measured before and after IFN-γ treatment. (E) Co-immunoprecipitation (co-IP) was used to detect endogenous interactions between AF6 and IRF1 in IECs. (F) 293T cells were co-transfected with constructs encoding hemagglutinin-tagged AF6 (HA-AF6) and Flag peptide-tagged IRF1 (Flag-IRF1); their interaction domains were detected by co-IP. (G) Immunoblotting was used to assess the expression of IRF1 and proteins encoded by its downstream genes in HT29 cells with or without IFN-γ exposure. (H) HT29 cells were transfected with constructs encoding AF6 with no or 3 nuclear localization signaling (NLS) domains (ΔNLS-AF6 and 3×NLS-AF6, respectively), and the expression and localization of IRF1 and proteins encoded by its downstream genes were assessed by immunoblotting following the separation of the nuclear and cytoplasmic fractions. Data are expressed as mean ± SEM. Pairwise comparisons between groups were conducted using two-tailed non-paired Student’s t tests. p -values were shown in the panel, and p < 0.05 indicates a significant difference.

Article Snippet: Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb , Cell Signaling Technology , Cat#9167.

Techniques: Expressing, Western Blot, Generated, Immunoprecipitation, Co-Immunoprecipitation Assay, Transfection, Construct, Two Tailed Test