foxp1 primary antibody  (Boster Bio)

Journal: Journal of Cellular and Molecular Medicine

Article Title: An integrated analysis of the anticarcinogenic role of forkhead box protein 1 in oesophageal squamous cell carcinoma

doi: 10.1111/jcmm.18294

Figure Lengend Snippet: The expression of forkhead box protein 1 (FOXP1) in oesophageal cancer and various tumours. (A) The expression of FOXP1 in various tumours in GEPIA database. (B) The expression of FOXP1 in oesophageal cancer tissues and normal tissues in UALCAN database. (C–E) The expression of FOXP1 in oesophageal cancer tissues depending on pathological type, tumour grade and tumour stage. (F–I) The expression of FOXP1 in oesophageal squamous cell carcinoma in GSE17351, GSE70409, GSE77861 and GSE100942 datasets. (J–K) The expression of FOXP1 in oesophageal squamous cell carcinoma tissues and adjacent normal tissues. * p < 0.05.

Article Snippet: FOXP1 primary antibody (1:150 dilution, BOSTER, M00723‐4) was added and incubated at room temperature for 1 h. The slices were incubated with the secondary antibody for 30 min at room temperature after washing with PBS buffer.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: An integrated analysis of the anticarcinogenic role of forkhead box protein 1 in oesophageal squamous cell carcinoma

doi: 10.1111/jcmm.18294

Figure Lengend Snippet: High expression of forkhead box protein 1 (FOXP1) indicates a better survival prognosis in oesophageal cancer. (A) The correlation between expression of FOXP1 and overall survival prognosis in different tumours in TISCH2 database. (B) Kaplan–Meier survival curves of overall survival for oesophageal cancer patients with high and low expression of FOXP1 in GEPIA database. (C) Kaplan–Meier survival curves of disease free survival for oesophageal cancer patients with high and low expression of FOXP1 in GEPIA database. (D) Kaplan–Meier survival curves of overall survival for oesophageal cancer patients with high and low expression of FOXP1 in KM plotter database. (E) Kaplan–Meier survival curves of disease free survival for oesophageal cancer patients with high and low expression of FOXP1 in KM plotter database. (F) Kaplan–Meier survival curves of overall survival for oesophageal squamous cell carcinoma patients with high and low expression of FOXP1 in KM plotter database. (G) Kaplan–Meier survival curves of overall survival for oesophageal squamous cell carcinoma patients with high and low expression of FOXP1 in GSE53625 dataset.

Article Snippet: FOXP1 primary antibody (1:150 dilution, BOSTER, M00723‐4) was added and incubated at room temperature for 1 h. The slices were incubated with the secondary antibody for 30 min at room temperature after washing with PBS buffer.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: An integrated analysis of the anticarcinogenic role of forkhead box protein 1 in oesophageal squamous cell carcinoma

doi: 10.1111/jcmm.18294

Figure Lengend Snippet: Identification of differential expression genes in high and low expression of forkhead box protein 1 (FOXP1) cohorts in oesophageal squamous cell carcinoma. (A) Venn diagram showing the intersection of differential expression genes in GSE26886 and GSE45168 datasets. (B, C) Volcano plots showing the differential expression genes in GSE26886 and GSE45168 datasets. (D, E) Cluster heatmaps of genes in GSE26886 and GSE45168 datasets.

Article Snippet: FOXP1 primary antibody (1:150 dilution, BOSTER, M00723‐4) was added and incubated at room temperature for 1 h. The slices were incubated with the secondary antibody for 30 min at room temperature after washing with PBS buffer.

Techniques: Quantitative Proteomics, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: An integrated analysis of the anticarcinogenic role of forkhead box protein 1 in oesophageal squamous cell carcinoma

doi: 10.1111/jcmm.18294

Figure Lengend Snippet: Forkhead box protein 1 (FOXP1) functional analysis and interaction network construction. (A) Gene ontology enrichment analysis for differential expression genes. (B) KEGG enrichment analysis for differential expression genes. (C) Network of enriched terms in metascape database. (D) The cord diagram of the correspondence between differential expression genes and signalling pathways. (E, F) Protein–protein interaction (PPI) network and MCODE components identified in differential expression genes in Metascape database. (G) The gene co‐expression network for FOXP1 in GeneMANIA database. (H) PPI network of differential expression genes in STRING database.

Article Snippet: FOXP1 primary antibody (1:150 dilution, BOSTER, M00723‐4) was added and incubated at room temperature for 1 h. The slices were incubated with the secondary antibody for 30 min at room temperature after washing with PBS buffer.

Techniques: Functional Assay, Quantitative Proteomics, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: An integrated analysis of the anticarcinogenic role of forkhead box protein 1 in oesophageal squamous cell carcinoma

doi: 10.1111/jcmm.18294

Figure Lengend Snippet: Association of forkhead box protein 1 (FOXP1) with the tumour immune microenvironment in oesophageal cancer. (A) The association of FOXP1 expression and tumour immune infiltration cells level in oesophageal cancer in TIMER database. (B) The relative proportion of different somatic copy number alterations (SCNA) states of FOXP1 for various TCGA tumour types in TIMER2.0 database. (C) The relationship between different SCNA states of FOXP1 and infiltration level of six types of immune cells in oesophageal cancer. * p < 0.05. (D) The relative proportion of the infiltration levels of 22 immune infiltrating cells in oesophageal squamous cell carcinoma based on GSE75241 dataset. (E) The immune cells correlation heatmap in oesophageal squamous cell carcinoma based on GSE75241 dataset.

Article Snippet: FOXP1 primary antibody (1:150 dilution, BOSTER, M00723‐4) was added and incubated at room temperature for 1 h. The slices were incubated with the secondary antibody for 30 min at room temperature after washing with PBS buffer.

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: An integrated analysis of the anticarcinogenic role of forkhead box protein 1 in oesophageal squamous cell carcinoma

doi: 10.1111/jcmm.18294

Figure Lengend Snippet: Forkhead box protein 1 (FOXP1) inhibits proliferation of oesophageal squamous cell carcinoma cells. (A, B) Western blot assay confirmed the expression levels of FOXP1 protein in different oesophageal squamous cell carcinoma cell lines and oesophageal epithelial cells. * p < 0.05, ** p < 0.01, ns p > 0.05. (C, D) Western blot assay confirmed the transfection efficiency of overexpression of FOXP1 in EC109 cells. ** p < 0.01. (E) The CCK‐8 assay showed overexpression of FOXP1 inhibits the proliferation of EC109 cells. *** p < 0.001. (F, G) The colony formation assay showed overexpression of FOXP1 inhibits the colony forming ability of EC109 cells. (H, I) The subcutaneous tumour formation assay showed overexpression of FOXP1 inhibits the ability of tumorigenicity of EC109 cells in vivo. * p < 0.05, ** p < 0.01, ns p > 0.05.

Article Snippet: FOXP1 primary antibody (1:150 dilution, BOSTER, M00723‐4) was added and incubated at room temperature for 1 h. The slices were incubated with the secondary antibody for 30 min at room temperature after washing with PBS buffer.

Techniques: Western Blot, Expressing, Transfection, Over Expression, CCK-8 Assay, Colony Assay, Tube Formation Assay, In Vivo

Journal: Nature Communications

Article Title: Foxp1 controls brown/beige adipocyte differentiation and thermogenesis through regulating β3-AR desensitization

doi: 10.1038/s41467-019-12988-8

Figure Lengend Snippet: Foxp1 expression in adipocytes is induced by adrenergic stimuli. a H&E staining and immunofluorescence (IF) analysis for the Foxp1 expression in BAT and sWAT from wild type mice at age of 4 weeks. DAPI, blue staining for nucleus; green color for Foxp1 expression. Bar, 50 μm. b Western blotting showed the four isoforms (A, B, D, C) of Foxp1 protein in BAT and sWAT from wild type mice at age of 2 months. c IHC analysis of FOXP1 expression in biopsies from PHEO patients and normal controls. Bar, 10 μm. d qPCR analysis of expression of Foxp1 and brown adipocyte markers ( C/ebpα , Pparγ , and Ucp1 ) during the time course of brown adipocyte differentiation from SVFs. e qPCR analysis of Foxp1 and β3-AR expression in BAT in mice with overnight 4 °C cold exposure. n = 3 biologically independent samples. f Western blotting of Foxp1 in BAT from mice above ( e ). g , h qPCR analysis of Foxp1 and β3-AR expression in brown adipocytes differentiated from murine ( g ) and human SVF ( h ) during an 8-hour CL-316,243 (0.1 μM) treatment as indicated. n = 3 biologically independent experiments. i Foxp1 expression profile in adipocytes derived from 3T3-L1 cells during an 8-h time course, stimulated by CL-316,243 (0.5 μM) with or without SB202190 (p38 kinase inhibitor, 10 μM), FR180204 (Erk1/2 inhibitors, 1 μM) and SCH772984 (Erk1/2 inhibitors, 10 μM), respectively. n = 3 biologically independent experiments. j Western blotting for Foxp1 in brown adipocytes derived from SVF, which were stimulated by CL-316,243 (0.1 μM) with or without SCH772984 (10 μM) for 8 h. * P < 0.05; ** P < 0.01; *** P < 0.001; error bar, mean ± SEM

Article Snippet: For immunofluorescence, heat-induced antigen retrieval with sodium citrate buffer (10 mM sodium citrate, 0.05% Tween 20, pH 6.0) was performed before bone sections were blocked with 10% normal serum containing 1% BSA in TBST (pH 7.6) for 2 h at room temperature, then incubated overnight at 4 °C with primary antibodies to mouse Foxp1 (Millipore, ABE68, 1:100), Ucp1 (Abcam, ab10893, 1:50), Prdm16 (Abcam, ab106410, 1:50).

Techniques: Expressing, Staining, Immunofluorescence, Western Blot, Derivative Assay

Journal: Nature Communications

Article Title: Foxp1 controls brown/beige adipocyte differentiation and thermogenesis through regulating β3-AR desensitization

doi: 10.1038/s41467-019-12988-8

Figure Lengend Snippet: Foxp1 deficiency promotes brown and beige adipocyte activation. a Representative dorsal view of BAT depot in Foxp1 fl/fl and Foxp1 Myf5 ∆/∆ mice at postnatal day 7. Bar, 1 cm. b HE (upper panel) and IHC staining (lower panel) with UCP1 antibody for BAT in ( a ). Bar, 10 μm. c , d qPCR analysis of Foxp1 and BAT-selective markers for BAT depot. n = 5 biologically independent samples. e Representative view of 3-month-old Foxp1 fl/fl and Foxp1 Ad ∆/∆ mice. Bar, 2 cm. f Representative view of BAT (upper panel) and sWAT depot (lower panel) in ( e ). g HE staining for BAT and sWAT in mutant mice. Bar, 50 μm. h IHC analysis with UCP1 antibody for BAT and sWAT upon 6-h cold exposure. Bar, 50 μm. i – k qPCR for expression of Foxp1 and BAT-selective genes in BAT and sWAT from Foxp1 fl/fl and Foxp1 Ad ∆/∆ mutant mice. n = 4 biologically independent mice/each group. * P < 0.05; ** P < 0.01; *** P < 0.001; error bar, mean ± SEM

Article Snippet: For immunofluorescence, heat-induced antigen retrieval with sodium citrate buffer (10 mM sodium citrate, 0.05% Tween 20, pH 6.0) was performed before bone sections were blocked with 10% normal serum containing 1% BSA in TBST (pH 7.6) for 2 h at room temperature, then incubated overnight at 4 °C with primary antibodies to mouse Foxp1 (Millipore, ABE68, 1:100), Ucp1 (Abcam, ab10893, 1:50), Prdm16 (Abcam, ab106410, 1:50).

Techniques: Activation Assay, Immunohistochemistry, Staining, Mutagenesis, Expressing

Journal: Nature Communications

Article Title: Foxp1 controls brown/beige adipocyte differentiation and thermogenesis through regulating β3-AR desensitization

doi: 10.1038/s41467-019-12988-8

Figure Lengend Snippet: Foxp1 deletion elevates energy expenditure and resists to HFD-induced obesity. a , b VO 2 and VCO 2 of Foxp1 fl/fl and Foxp1 Ad ∆/∆ mice in metabolic cages at age of 3 months old. n = 7 biologically independent mice/each group. c , d Quantification of O 2 and CO 2 consumption in light and dark. e Dorsal view of Foxp1 fl/fl and Foxp1 Ad ∆/∆ mice after 8-week feeding with HFD at age of 6 months old. Bar, 2 cm. f Representative depot of BAT and sWAT in HFD-fed mice ( e ). Bar, 1 cm. g Body weight of HFD-fed mice. h Relative adiposity of HFD-fed mice. i Statistics of rectal temperature of HFD-fed mice. j , k GTT and ITT of HFD-fed mice. * P < 0.05; ** P < 0.01; *** P < 0.001; n = 7 biologically independent mice/each group; error bar, mean ± SEM

Article Snippet: For immunofluorescence, heat-induced antigen retrieval with sodium citrate buffer (10 mM sodium citrate, 0.05% Tween 20, pH 6.0) was performed before bone sections were blocked with 10% normal serum containing 1% BSA in TBST (pH 7.6) for 2 h at room temperature, then incubated overnight at 4 °C with primary antibodies to mouse Foxp1 (Millipore, ABE68, 1:100), Ucp1 (Abcam, ab10893, 1:50), Prdm16 (Abcam, ab106410, 1:50).

Techniques:

Journal: Nature Communications

Article Title: Foxp1 controls brown/beige adipocyte differentiation and thermogenesis through regulating β3-AR desensitization

doi: 10.1038/s41467-019-12988-8

Figure Lengend Snippet: Overexpression of Foxp1 represses adaptive thermogenesis and is sensitive to HFD-induce obesity. a Representative bioluminescence imaging of two stains of aP2-Foxp1;Ucp1-Luc transgenic mice after 6-hour cold exposure. n = 3 biologically independent mice/each group. Bar, 2 cm. b Representative picture of BAT depot in aP2-Foxp1 transgenic mice. Bar, 0.5 cm. c H&E staining for adipose sections of BAT and sWAT from transgenic mice in ( b ). Bar, 10 μm. d IHC with UCP1 antibody for adipose sections from transgenic mice after 6-h cold exposure. Bar, 10 μm. e Intracellular structure of adipocytes in BAT and sWAT from aP2-Foxp1 mice, as showed by transmission electronic microscope (TEM). Bar, 2 μm. f Assessment of mitochondria DNA abundance by qPCR for the BAT and sWAT from transgenic mice. n = 4 biologically independent mice/each group. g qPCR analysis for Ucp1 expression in adipose tissues of transgenic mice. n = 4 biologically independent samples. h Western blotting for UCP1, HSL, and phosphorylated HSL in BAT and sWAT. i Representative dorsal view of aP2-Foxp1 transgenic mice subjected to 8-week HFD feeding since at age of 3 months. Bar, 2 cm. j Relative adiposity in transgenic mice ( i ). n = 4 biologically independent mice/each group. k The heat map of relative expression of adipogenic genes in three strains of aP2-Foxp1 transgenic mice at age of 3 weeks, as evaluated by RNA-seq. l qPCR analysis validated the BAT-selective genes expression in BAT from ( k ). n = 3 biologically independent mice/each group; * P < 0.05; ** P < 0.01; *** P < 0.001; error bar, mean ± SEM

Article Snippet: For immunofluorescence, heat-induced antigen retrieval with sodium citrate buffer (10 mM sodium citrate, 0.05% Tween 20, pH 6.0) was performed before bone sections were blocked with 10% normal serum containing 1% BSA in TBST (pH 7.6) for 2 h at room temperature, then incubated overnight at 4 °C with primary antibodies to mouse Foxp1 (Millipore, ABE68, 1:100), Ucp1 (Abcam, ab10893, 1:50), Prdm16 (Abcam, ab106410, 1:50).

Techniques: Over Expression, Imaging, Transgenic Assay, Staining, Transmission Assay, Microscopy, Expressing, Western Blot, RNA Sequencing Assay

Journal: Nature Communications

Article Title: Foxp1 controls brown/beige adipocyte differentiation and thermogenesis through regulating β3-AR desensitization

doi: 10.1038/s41467-019-12988-8

Figure Lengend Snippet: Foxp1 represses β3-AR transcription in adipocytes. a, b β3-AR expression in BAT and sWAT were assessed by qPCR and western blotting in aP2-Foxp1 transgenic mice at 2 month old. n = 3 biologically independent samples/each group. c, d β3-AR expression in BAT and sWAT were assessed by qPCR and western blotting in Foxp1 aP2 ∆/∆ knockout mice at 2 months old. n = 3 biologically independent samples/each group. e ChIP-seq profile showed the Foxp1 binding sites within Adrb3 gene promoter region. Red arrows indicated the potential Foxp1 binding sites. Adrb3 gene was reversely transcribed. ChIP was based on SVF cells derived from BAT. f Upper panel showed the location of Foxp1 binding site at −2251 bp upstream of β3-AR gene promoter with a schematic drawing. ChIP-PCR confirmed the relative enrichment of Foxp1 binding sites (lower panel). n = 3 biologically independent samples/each group. g Basic OCR of SVF-derived brown adipocytes from Foxp1 aP2 ∆/∆ mice, which were transfected β3-AR -shRNA lentivirus or administrated with β3-AR inhibitor L748337 (10 μM) for 1 h. n = 3 biologically independent experiments. h Representative infrared imaging of aP2-Foxp1 transgenic mice with or without 6-hour CL-316,243 (10 μM) exposure. Bar, 1 cm. i Quantification of average skin temperature in boxed regions in ( h ). n = 4 biologically independent mice/each group. Samples were isolated from mice at age of 2 months. j CL-stimulated β3-AR desensitization in SVF-derived brown adipocytes from aP2-Foxp1 transgenic mice and Foxp1 aP2 ∆/∆ knockout mice. β3-AR expression was assessed by qPCR during the 6-h time course of CL-316243 (0.1 μM) treatment. n = 3 biologically independent samples/each group. * P < 0.05; ** P < 0.01; *** P < 0.001; error bar, mean ± SEM

Article Snippet: For immunofluorescence, heat-induced antigen retrieval with sodium citrate buffer (10 mM sodium citrate, 0.05% Tween 20, pH 6.0) was performed before bone sections were blocked with 10% normal serum containing 1% BSA in TBST (pH 7.6) for 2 h at room temperature, then incubated overnight at 4 °C with primary antibodies to mouse Foxp1 (Millipore, ABE68, 1:100), Ucp1 (Abcam, ab10893, 1:50), Prdm16 (Abcam, ab106410, 1:50).

Techniques: Expressing, Western Blot, Transgenic Assay, Knock-Out, ChIP-sequencing, Binding Assay, Derivative Assay, Transfection, shRNA, Imaging, Isolation

Journal: Nature Communications

Article Title: Foxp1 controls brown/beige adipocyte differentiation and thermogenesis through regulating β3-AR desensitization

doi: 10.1038/s41467-019-12988-8

Figure Lengend Snippet: Foxp1 physically interacts with Prdm16-C/ebpβ complex in brown adipocytes. a , b Co-immunoprecipitation validated the physical interaction between Foxp1, C/ebpβ and Prdm16 proteins in cell lysates from 293T cell lines transfected with indicated plasmids ( a ), or brown adipocytes induced from stromal vascular fractions ( b ). c Immunofluorescence analysis showed the co-localization of Foxp1 and Prdm16 in the nucleus of BAT from wild type mice at age of 1 month old. Bar, 10 μm. d Luciferase reporter assay showed the transactivation of PPARγ -Luc by Foxp1 and Prdm16 protein in 3T3-L1 cell lines. n = 3 biologically independent samples/each group. e Luciferase reporter assay showed the transactivation of β3-AR -Luc by Foxp1, C/ebpβ and Prdm16 protein in 293T cell lines. n = 3 biologically independent experiments; * P < 0.05, ** P < 0.01, *** P < 0.001; error bar, mean ± SEM. f Diagram depicting the mechanisms that Foxp1 represses brown/beige adipocyte differentiation and adaptive thermogenesis. Expression of Foxp1 is induced by β3-AR/cAMP/Erk1/2 cascades. Conversely, Foxp1 suppresses β3-AR transcription by counteracting the activity of Prdm16-C/ebpβ complex

Article Snippet: For immunofluorescence, heat-induced antigen retrieval with sodium citrate buffer (10 mM sodium citrate, 0.05% Tween 20, pH 6.0) was performed before bone sections were blocked with 10% normal serum containing 1% BSA in TBST (pH 7.6) for 2 h at room temperature, then incubated overnight at 4 °C with primary antibodies to mouse Foxp1 (Millipore, ABE68, 1:100), Ucp1 (Abcam, ab10893, 1:50), Prdm16 (Abcam, ab106410, 1:50).

Techniques: Immunoprecipitation, Transfection, Immunofluorescence, Luciferase, Reporter Assay, Expressing, Activity Assay