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Structured Review

Proteintech anti fabp5 antibody
a , Experimental schematic of untargeted metabolomic profiling on the culture supernatant of IECs isolated from Tm6sf2 ΔIEC and Tm6sf2 fl mice. Created with BioRender.com . b , PCA analysis and heat map of differential metabolites secreted by IECs from mice ( n = 8 per group). c , Metabolomics targeting free fatty acids on the cultural supernatant of isolated IECs ( n = 8 per group). d , Representative small intestine images of H&E staining and intestinal triglyceride of Tm6sf2 ΔIEC mice fed with NC ( n = 5 per group) or CD-HFD ( n = 9 per group). e , Silver staining coupled with mass spectrometry analysis after pull-down assay on mouse intestinal tissues. f , Co-immunoprecipitation of TM6SF2 and <t>FABP5</t> using mouse intestinal proteins. g , MST assay for direct binding between TM6SF2 and FABP5 with dissociation constant ( K d ) provided. h , Representative structure of molecular docking between TM6SF2 (purple) and FABP5 (blue). i , Metabolomics targeting free fatty acids in TM6SF2 KO Caco2 cells with or without FABP5 overexpression ( n = 6 per group). j , Experimental schematic and intestinal permeability by FITC-labelled dextran assay in mice supplemented with the top differential free fatty acids ( n = 5 per group). k , Permeability of Caco2 cell monolayers treated with arachidic acid or arachidonic acid ( n = 3 per group). l , m , Heat map of differential faecal microorganisms ( l ) and LPA-targeting metabolomics on portal vein serum ( m ) of mice treated with free fatty acids ( n = 4–5 per group). Results are presented as the mean ± s.d. Experiments were repeated three times with similar results ( e and f ). Statistical significance was determined by two-tailed Student’s t -test ( c , d , i and k ) or one-way ANOVA followed by Turkey’s multiple comparison ( j and m ).
Anti Fabp5 Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Images

1) Product Images from "Intestinal TM6SF2 protects against metabolic dysfunction-associated steatohepatitis through the gut–liver axis"

Article Title: Intestinal TM6SF2 protects against metabolic dysfunction-associated steatohepatitis through the gut–liver axis

Journal: Nature Metabolism

doi: 10.1038/s42255-024-01177-7

a , Experimental schematic of untargeted metabolomic profiling on the culture supernatant of IECs isolated from Tm6sf2 ΔIEC and Tm6sf2 fl mice. Created with BioRender.com . b , PCA analysis and heat map of differential metabolites secreted by IECs from mice ( n = 8 per group). c , Metabolomics targeting free fatty acids on the cultural supernatant of isolated IECs ( n = 8 per group). d , Representative small intestine images of H&E staining and intestinal triglyceride of Tm6sf2 ΔIEC mice fed with NC ( n = 5 per group) or CD-HFD ( n = 9 per group). e , Silver staining coupled with mass spectrometry analysis after pull-down assay on mouse intestinal tissues. f , Co-immunoprecipitation of TM6SF2 and FABP5 using mouse intestinal proteins. g , MST assay for direct binding between TM6SF2 and FABP5 with dissociation constant ( K d ) provided. h , Representative structure of molecular docking between TM6SF2 (purple) and FABP5 (blue). i , Metabolomics targeting free fatty acids in TM6SF2 KO Caco2 cells with or without FABP5 overexpression ( n = 6 per group). j , Experimental schematic and intestinal permeability by FITC-labelled dextran assay in mice supplemented with the top differential free fatty acids ( n = 5 per group). k , Permeability of Caco2 cell monolayers treated with arachidic acid or arachidonic acid ( n = 3 per group). l , m , Heat map of differential faecal microorganisms ( l ) and LPA-targeting metabolomics on portal vein serum ( m ) of mice treated with free fatty acids ( n = 4–5 per group). Results are presented as the mean ± s.d. Experiments were repeated three times with similar results ( e and f ). Statistical significance was determined by two-tailed Student’s t -test ( c , d , i and k ) or one-way ANOVA followed by Turkey’s multiple comparison ( j and m ).
Figure Legend Snippet: a , Experimental schematic of untargeted metabolomic profiling on the culture supernatant of IECs isolated from Tm6sf2 ΔIEC and Tm6sf2 fl mice. Created with BioRender.com . b , PCA analysis and heat map of differential metabolites secreted by IECs from mice ( n = 8 per group). c , Metabolomics targeting free fatty acids on the cultural supernatant of isolated IECs ( n = 8 per group). d , Representative small intestine images of H&E staining and intestinal triglyceride of Tm6sf2 ΔIEC mice fed with NC ( n = 5 per group) or CD-HFD ( n = 9 per group). e , Silver staining coupled with mass spectrometry analysis after pull-down assay on mouse intestinal tissues. f , Co-immunoprecipitation of TM6SF2 and FABP5 using mouse intestinal proteins. g , MST assay for direct binding between TM6SF2 and FABP5 with dissociation constant ( K d ) provided. h , Representative structure of molecular docking between TM6SF2 (purple) and FABP5 (blue). i , Metabolomics targeting free fatty acids in TM6SF2 KO Caco2 cells with or without FABP5 overexpression ( n = 6 per group). j , Experimental schematic and intestinal permeability by FITC-labelled dextran assay in mice supplemented with the top differential free fatty acids ( n = 5 per group). k , Permeability of Caco2 cell monolayers treated with arachidic acid or arachidonic acid ( n = 3 per group). l , m , Heat map of differential faecal microorganisms ( l ) and LPA-targeting metabolomics on portal vein serum ( m ) of mice treated with free fatty acids ( n = 4–5 per group). Results are presented as the mean ± s.d. Experiments were repeated three times with similar results ( e and f ). Statistical significance was determined by two-tailed Student’s t -test ( c , d , i and k ) or one-way ANOVA followed by Turkey’s multiple comparison ( j and m ).

Techniques Used: Isolation, Staining, Silver Staining, Mass Spectrometry, Pull Down Assay, Immunoprecipitation, Binding Assay, Over Expression, Permeability, Two Tailed Test, Comparison



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a , Experimental schematic of untargeted metabolomic profiling on the culture supernatant of IECs isolated from Tm6sf2 ΔIEC and Tm6sf2 fl mice. Created with BioRender.com . b , PCA analysis and heat map of differential metabolites secreted by IECs from mice ( n = 8 per group). c , Metabolomics targeting free fatty acids on the cultural supernatant of isolated IECs ( n = 8 per group). d , Representative small intestine images of H&E staining and intestinal triglyceride of Tm6sf2 ΔIEC mice fed with NC ( n = 5 per group) or CD-HFD ( n = 9 per group). e , Silver staining coupled with mass spectrometry analysis after pull-down assay on mouse intestinal tissues. f , Co-immunoprecipitation of TM6SF2 and <t>FABP5</t> using mouse intestinal proteins. g , MST assay for direct binding between TM6SF2 and FABP5 with dissociation constant ( K d ) provided. h , Representative structure of molecular docking between TM6SF2 (purple) and FABP5 (blue). i , Metabolomics targeting free fatty acids in TM6SF2 KO Caco2 cells with or without FABP5 overexpression ( n = 6 per group). j , Experimental schematic and intestinal permeability by FITC-labelled dextran assay in mice supplemented with the top differential free fatty acids ( n = 5 per group). k , Permeability of Caco2 cell monolayers treated with arachidic acid or arachidonic acid ( n = 3 per group). l , m , Heat map of differential faecal microorganisms ( l ) and LPA-targeting metabolomics on portal vein serum ( m ) of mice treated with free fatty acids ( n = 4–5 per group). Results are presented as the mean ± s.d. Experiments were repeated three times with similar results ( e and f ). Statistical significance was determined by two-tailed Student’s t -test ( c , d , i and k ) or one-way ANOVA followed by Turkey’s multiple comparison ( j and m ).
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a , Experimental schematic of untargeted metabolomic profiling on the culture supernatant of IECs isolated from Tm6sf2 ΔIEC and Tm6sf2 fl mice. Created with BioRender.com . b , PCA analysis and heat map of differential metabolites secreted by IECs from mice ( n = 8 per group). c , Metabolomics targeting free fatty acids on the cultural supernatant of isolated IECs ( n = 8 per group). d , Representative small intestine images of H&E staining and intestinal triglyceride of Tm6sf2 ΔIEC mice fed with NC ( n = 5 per group) or CD-HFD ( n = 9 per group). e , Silver staining coupled with mass spectrometry analysis after pull-down assay on mouse intestinal tissues. f , Co-immunoprecipitation of TM6SF2 and <t>FABP5</t> using mouse intestinal proteins. g , MST assay for direct binding between TM6SF2 and FABP5 with dissociation constant ( K d ) provided. h , Representative structure of molecular docking between TM6SF2 (purple) and FABP5 (blue). i , Metabolomics targeting free fatty acids in TM6SF2 KO Caco2 cells with or without FABP5 overexpression ( n = 6 per group). j , Experimental schematic and intestinal permeability by FITC-labelled dextran assay in mice supplemented with the top differential free fatty acids ( n = 5 per group). k , Permeability of Caco2 cell monolayers treated with arachidic acid or arachidonic acid ( n = 3 per group). l , m , Heat map of differential faecal microorganisms ( l ) and LPA-targeting metabolomics on portal vein serum ( m ) of mice treated with free fatty acids ( n = 4–5 per group). Results are presented as the mean ± s.d. Experiments were repeated three times with similar results ( e and f ). Statistical significance was determined by two-tailed Student’s t -test ( c , d , i and k ) or one-way ANOVA followed by Turkey’s multiple comparison ( j and m ).
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The expression of specific transcription factors, membrane-associated proteins and cell markers in human small and large intestine. A Immunoblot of protein FOXM1, MYBL2 and UBE2C in villus and crypt derived from mouse small intestine. B FOXM1 and KI67 immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. C UBE2C immunostaining of large intestine derived from human specimens. Scale bars: 10 μm. D MYBL2 and KI67 immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. E Immunoblot of protein ANPEP, SLC12A2, IGF1R and SLC16A1 in villus and crypt derived from mouse small intestine. F SLC12A2 immunostaining of small intestine derived from human specimens. Scale bars: 50 μm. G SLC16A1 and E-Cadherin immunostaining of small intestine derived from human specimens. Scale bars: 50 μm. H ANPEP and E-Cadherin immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. I IGF1R immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. J Immunoblot of protein HSPD1, CPE, <t>FABP5</t> and C1QBP in villus and crypt derived from mouse small intestine. K CPE and Chromogranin A immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. L HSPD1 and E-Cadherin immunostaining of small intestine derived from human specimens. Scale bars: 50 μm. M FABP5 and Chromogranin A immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. N C1QBP and E-Cadherin immunostaining of small intestine derived from human specimens. Scale bars: 50 μm
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Image Search Results


a , Experimental schematic of untargeted metabolomic profiling on the culture supernatant of IECs isolated from Tm6sf2 ΔIEC and Tm6sf2 fl mice. Created with BioRender.com . b , PCA analysis and heat map of differential metabolites secreted by IECs from mice ( n = 8 per group). c , Metabolomics targeting free fatty acids on the cultural supernatant of isolated IECs ( n = 8 per group). d , Representative small intestine images of H&E staining and intestinal triglyceride of Tm6sf2 ΔIEC mice fed with NC ( n = 5 per group) or CD-HFD ( n = 9 per group). e , Silver staining coupled with mass spectrometry analysis after pull-down assay on mouse intestinal tissues. f , Co-immunoprecipitation of TM6SF2 and FABP5 using mouse intestinal proteins. g , MST assay for direct binding between TM6SF2 and FABP5 with dissociation constant ( K d ) provided. h , Representative structure of molecular docking between TM6SF2 (purple) and FABP5 (blue). i , Metabolomics targeting free fatty acids in TM6SF2 KO Caco2 cells with or without FABP5 overexpression ( n = 6 per group). j , Experimental schematic and intestinal permeability by FITC-labelled dextran assay in mice supplemented with the top differential free fatty acids ( n = 5 per group). k , Permeability of Caco2 cell monolayers treated with arachidic acid or arachidonic acid ( n = 3 per group). l , m , Heat map of differential faecal microorganisms ( l ) and LPA-targeting metabolomics on portal vein serum ( m ) of mice treated with free fatty acids ( n = 4–5 per group). Results are presented as the mean ± s.d. Experiments were repeated three times with similar results ( e and f ). Statistical significance was determined by two-tailed Student’s t -test ( c , d , i and k ) or one-way ANOVA followed by Turkey’s multiple comparison ( j and m ).

Journal: Nature Metabolism

Article Title: Intestinal TM6SF2 protects against metabolic dysfunction-associated steatohepatitis through the gut–liver axis

doi: 10.1038/s42255-024-01177-7

Figure Lengend Snippet: a , Experimental schematic of untargeted metabolomic profiling on the culture supernatant of IECs isolated from Tm6sf2 ΔIEC and Tm6sf2 fl mice. Created with BioRender.com . b , PCA analysis and heat map of differential metabolites secreted by IECs from mice ( n = 8 per group). c , Metabolomics targeting free fatty acids on the cultural supernatant of isolated IECs ( n = 8 per group). d , Representative small intestine images of H&E staining and intestinal triglyceride of Tm6sf2 ΔIEC mice fed with NC ( n = 5 per group) or CD-HFD ( n = 9 per group). e , Silver staining coupled with mass spectrometry analysis after pull-down assay on mouse intestinal tissues. f , Co-immunoprecipitation of TM6SF2 and FABP5 using mouse intestinal proteins. g , MST assay for direct binding between TM6SF2 and FABP5 with dissociation constant ( K d ) provided. h , Representative structure of molecular docking between TM6SF2 (purple) and FABP5 (blue). i , Metabolomics targeting free fatty acids in TM6SF2 KO Caco2 cells with or without FABP5 overexpression ( n = 6 per group). j , Experimental schematic and intestinal permeability by FITC-labelled dextran assay in mice supplemented with the top differential free fatty acids ( n = 5 per group). k , Permeability of Caco2 cell monolayers treated with arachidic acid or arachidonic acid ( n = 3 per group). l , m , Heat map of differential faecal microorganisms ( l ) and LPA-targeting metabolomics on portal vein serum ( m ) of mice treated with free fatty acids ( n = 4–5 per group). Results are presented as the mean ± s.d. Experiments were repeated three times with similar results ( e and f ). Statistical significance was determined by two-tailed Student’s t -test ( c , d , i and k ) or one-way ANOVA followed by Turkey’s multiple comparison ( j and m ).

Article Snippet: The protein mixture was also incubated overnight at 4 °C with either anti-TM6SF2 antibody or anti-FABP5 antibody (RRID: AB_2100341, 12348-1-AP, Proteintech).

Techniques: Isolation, Staining, Silver Staining, Mass Spectrometry, Pull Down Assay, Immunoprecipitation, Binding Assay, Over Expression, Permeability, Two Tailed Test, Comparison

Primers used for reverse transcription quantitative PCR.

Journal: Molecular Medicine Reports

Article Title: Stigmasterol exerts antiglioma effects by regulating lipid metabolism

doi: 10.3892/mmr.2024.13351

Figure Lengend Snippet: Primers used for reverse transcription quantitative PCR.

Article Snippet: The following primary antibodies were utilized for western blotting: Rabbit anti-fatty acid binding protein 5 (FABP5) antibody (1:2,000; cat. no. 12348-1-AP; Proteintech Group, Inc.), rabbit anti-α-1B adrenergic receptor (ADRA1B) antibody (1:1,000; cat. no. 22419-1-AP; Proteintech Group, Inc.) and mouse anti-β-actin antibody (1:5,000; cat. no. T0022; Affinity Biosciences).

Techniques: Reverse Transcription, Sequencing

Construction and validation of a prognostic model associated with stigmasterol target genes. (A) Venn diagram of potential targets of stigmasterol in the treatment of GBM, prognosis-related genes of patients with GBM and lipid metabolism-related genes. (B) Two prognosis-related target mRNAs of stigmasterol for GBM treatment were selected. (C and D) Least absolute shrinkage and selection operator variable screening process. (E) Distribution of survival status and risk score of patients with GBM in the training set. (F) Expression characteristics of the two prognosis-related target mRNAs of stigmasterol for GBM treatment in the training set. (G) ROC curve for patient survival of the training set. (H) Survival analysis between the two risk subgroups in the training set. (I) ROC curve for patient survival of the internal validation set. (J) Survival analysis between the two risk subgroups in the internal validation set. (K) ROC curve for patient survival of the external validation set. (L) Survival analysis between the two risk subgroups in the external validation set. GBM, glioblastoma; ROC, receiver operating characteristic; FABP5, fatty acid binding protein 5; ADRA1B, α-1B adrenergic receptor; AUC, area under the curve.

Journal: Molecular Medicine Reports

Article Title: Stigmasterol exerts antiglioma effects by regulating lipid metabolism

doi: 10.3892/mmr.2024.13351

Figure Lengend Snippet: Construction and validation of a prognostic model associated with stigmasterol target genes. (A) Venn diagram of potential targets of stigmasterol in the treatment of GBM, prognosis-related genes of patients with GBM and lipid metabolism-related genes. (B) Two prognosis-related target mRNAs of stigmasterol for GBM treatment were selected. (C and D) Least absolute shrinkage and selection operator variable screening process. (E) Distribution of survival status and risk score of patients with GBM in the training set. (F) Expression characteristics of the two prognosis-related target mRNAs of stigmasterol for GBM treatment in the training set. (G) ROC curve for patient survival of the training set. (H) Survival analysis between the two risk subgroups in the training set. (I) ROC curve for patient survival of the internal validation set. (J) Survival analysis between the two risk subgroups in the internal validation set. (K) ROC curve for patient survival of the external validation set. (L) Survival analysis between the two risk subgroups in the external validation set. GBM, glioblastoma; ROC, receiver operating characteristic; FABP5, fatty acid binding protein 5; ADRA1B, α-1B adrenergic receptor; AUC, area under the curve.

Article Snippet: The following primary antibodies were utilized for western blotting: Rabbit anti-fatty acid binding protein 5 (FABP5) antibody (1:2,000; cat. no. 12348-1-AP; Proteintech Group, Inc.), rabbit anti-α-1B adrenergic receptor (ADRA1B) antibody (1:1,000; cat. no. 22419-1-AP; Proteintech Group, Inc.) and mouse anti-β-actin antibody (1:5,000; cat. no. T0022; Affinity Biosciences).

Techniques: Selection, Expressing, Binding Assay

Effects of stigmasterol on lipid metabolism and the expression of related target genes in GBM cells. (A) Effect of stigmasterol on free fatty acid levels in GBM cells. (B) Effect of stigmasterol on total cholesterol levels in GBM cells. (C) Effect of stigmasterol on the mRNA expression of the two target genes (FABP5 and ADRA1B) in GBM cells. (D) Effect of stigmasterol on the expression of the proteins encoded by the two target genes in GBM cells. (E) Molecular docking of the protein encoded by FABP5 with stigmasterol. (F) Molecular docking of the protein encoded by ADRA1B with stigmasterol. *P<0.05 and **P<0.01. GBM, glioblastoma; FABP5, fatty acid binding protein 5; ADRA1B, α-1B adrenergic receptor.

Journal: Molecular Medicine Reports

Article Title: Stigmasterol exerts antiglioma effects by regulating lipid metabolism

doi: 10.3892/mmr.2024.13351

Figure Lengend Snippet: Effects of stigmasterol on lipid metabolism and the expression of related target genes in GBM cells. (A) Effect of stigmasterol on free fatty acid levels in GBM cells. (B) Effect of stigmasterol on total cholesterol levels in GBM cells. (C) Effect of stigmasterol on the mRNA expression of the two target genes (FABP5 and ADRA1B) in GBM cells. (D) Effect of stigmasterol on the expression of the proteins encoded by the two target genes in GBM cells. (E) Molecular docking of the protein encoded by FABP5 with stigmasterol. (F) Molecular docking of the protein encoded by ADRA1B with stigmasterol. *P<0.05 and **P<0.01. GBM, glioblastoma; FABP5, fatty acid binding protein 5; ADRA1B, α-1B adrenergic receptor.

Article Snippet: The following primary antibodies were utilized for western blotting: Rabbit anti-fatty acid binding protein 5 (FABP5) antibody (1:2,000; cat. no. 12348-1-AP; Proteintech Group, Inc.), rabbit anti-α-1B adrenergic receptor (ADRA1B) antibody (1:1,000; cat. no. 22419-1-AP; Proteintech Group, Inc.) and mouse anti-β-actin antibody (1:5,000; cat. no. T0022; Affinity Biosciences).

Techniques: Expressing, Binding Assay

Primers used for reverse transcription quantitative PCR.

Journal: Molecular Medicine Reports

Article Title: Stigmasterol exerts antiglioma effects by regulating lipid metabolism

doi: 10.3892/mmr.2024.13351

Figure Lengend Snippet: Primers used for reverse transcription quantitative PCR.

Article Snippet: For detection, the following secondary antibodies were used: HRP-conjugated goat anti-rabbit immunoglobulin G (IgG) antibody (dilution, 1:5,000; cat. no. A0208; Beyotime Institute of Biotechnology) for FABP5 and ADRA1B detection, and HRP-conjugated goat anti-mouse IgG antibody (dilution, 1:5,000, cat. no. SA00001-1; Proteintech Group, Inc.) for β-actin detection.

Techniques: Reverse Transcription, Sequencing

Construction and validation of a prognostic model associated with stigmasterol target genes. (A) Venn diagram of potential targets of stigmasterol in the treatment of GBM, prognosis-related genes of patients with GBM and lipid metabolism-related genes. (B) Two prognosis-related target mRNAs of stigmasterol for GBM treatment were selected. (C and D) Least absolute shrinkage and selection operator variable screening process. (E) Distribution of survival status and risk score of patients with GBM in the training set. (F) Expression characteristics of the two prognosis-related target mRNAs of stigmasterol for GBM treatment in the training set. (G) ROC curve for patient survival of the training set. (H) Survival analysis between the two risk subgroups in the training set. (I) ROC curve for patient survival of the internal validation set. (J) Survival analysis between the two risk subgroups in the internal validation set. (K) ROC curve for patient survival of the external validation set. (L) Survival analysis between the two risk subgroups in the external validation set. GBM, glioblastoma; ROC, receiver operating characteristic; FABP5, fatty acid binding protein 5; ADRA1B, α-1B adrenergic receptor; AUC, area under the curve.

Journal: Molecular Medicine Reports

Article Title: Stigmasterol exerts antiglioma effects by regulating lipid metabolism

doi: 10.3892/mmr.2024.13351

Figure Lengend Snippet: Construction and validation of a prognostic model associated with stigmasterol target genes. (A) Venn diagram of potential targets of stigmasterol in the treatment of GBM, prognosis-related genes of patients with GBM and lipid metabolism-related genes. (B) Two prognosis-related target mRNAs of stigmasterol for GBM treatment were selected. (C and D) Least absolute shrinkage and selection operator variable screening process. (E) Distribution of survival status and risk score of patients with GBM in the training set. (F) Expression characteristics of the two prognosis-related target mRNAs of stigmasterol for GBM treatment in the training set. (G) ROC curve for patient survival of the training set. (H) Survival analysis between the two risk subgroups in the training set. (I) ROC curve for patient survival of the internal validation set. (J) Survival analysis between the two risk subgroups in the internal validation set. (K) ROC curve for patient survival of the external validation set. (L) Survival analysis between the two risk subgroups in the external validation set. GBM, glioblastoma; ROC, receiver operating characteristic; FABP5, fatty acid binding protein 5; ADRA1B, α-1B adrenergic receptor; AUC, area under the curve.

Article Snippet: For detection, the following secondary antibodies were used: HRP-conjugated goat anti-rabbit immunoglobulin G (IgG) antibody (dilution, 1:5,000; cat. no. A0208; Beyotime Institute of Biotechnology) for FABP5 and ADRA1B detection, and HRP-conjugated goat anti-mouse IgG antibody (dilution, 1:5,000, cat. no. SA00001-1; Proteintech Group, Inc.) for β-actin detection.

Techniques: Selection, Expressing, Binding Assay

Effects of stigmasterol on lipid metabolism and the expression of related target genes in GBM cells. (A) Effect of stigmasterol on free fatty acid levels in GBM cells. (B) Effect of stigmasterol on total cholesterol levels in GBM cells. (C) Effect of stigmasterol on the mRNA expression of the two target genes (FABP5 and ADRA1B) in GBM cells. (D) Effect of stigmasterol on the expression of the proteins encoded by the two target genes in GBM cells. (E) Molecular docking of the protein encoded by FABP5 with stigmasterol. (F) Molecular docking of the protein encoded by ADRA1B with stigmasterol. *P<0.05 and **P<0.01. GBM, glioblastoma; FABP5, fatty acid binding protein 5; ADRA1B, α-1B adrenergic receptor.

Journal: Molecular Medicine Reports

Article Title: Stigmasterol exerts antiglioma effects by regulating lipid metabolism

doi: 10.3892/mmr.2024.13351

Figure Lengend Snippet: Effects of stigmasterol on lipid metabolism and the expression of related target genes in GBM cells. (A) Effect of stigmasterol on free fatty acid levels in GBM cells. (B) Effect of stigmasterol on total cholesterol levels in GBM cells. (C) Effect of stigmasterol on the mRNA expression of the two target genes (FABP5 and ADRA1B) in GBM cells. (D) Effect of stigmasterol on the expression of the proteins encoded by the two target genes in GBM cells. (E) Molecular docking of the protein encoded by FABP5 with stigmasterol. (F) Molecular docking of the protein encoded by ADRA1B with stigmasterol. *P<0.05 and **P<0.01. GBM, glioblastoma; FABP5, fatty acid binding protein 5; ADRA1B, α-1B adrenergic receptor.

Article Snippet: For detection, the following secondary antibodies were used: HRP-conjugated goat anti-rabbit immunoglobulin G (IgG) antibody (dilution, 1:5,000; cat. no. A0208; Beyotime Institute of Biotechnology) for FABP5 and ADRA1B detection, and HRP-conjugated goat anti-mouse IgG antibody (dilution, 1:5,000, cat. no. SA00001-1; Proteintech Group, Inc.) for β-actin detection.

Techniques: Expressing, Binding Assay

Primers used for reverse transcription quantitative PCR.

Journal: Molecular Medicine Reports

Article Title: Stigmasterol exerts antiglioma effects by regulating lipid metabolism

doi: 10.3892/mmr.2024.13351

Figure Lengend Snippet: Primers used for reverse transcription quantitative PCR.

Article Snippet: For detection, the following secondary antibodies were used: HRP-conjugated goat anti-rabbit immunoglobulin G (IgG) antibody (dilution, 1:5,000; cat. no. A0208; Beyotime Institute of Biotechnology) for FABP5 and ADRA1B detection, and HRP-conjugated goat anti-mouse IgG antibody (dilution, 1:5,000, cat. no. SA00001-1; Proteintech Group, Inc.) for β-actin detection.

Techniques: Reverse Transcription, Sequencing

Construction and validation of a prognostic model associated with stigmasterol target genes. (A) Venn diagram of potential targets of stigmasterol in the treatment of GBM, prognosis-related genes of patients with GBM and lipid metabolism-related genes. (B) Two prognosis-related target mRNAs of stigmasterol for GBM treatment were selected. (C and D) Least absolute shrinkage and selection operator variable screening process. (E) Distribution of survival status and risk score of patients with GBM in the training set. (F) Expression characteristics of the two prognosis-related target mRNAs of stigmasterol for GBM treatment in the training set. (G) ROC curve for patient survival of the training set. (H) Survival analysis between the two risk subgroups in the training set. (I) ROC curve for patient survival of the internal validation set. (J) Survival analysis between the two risk subgroups in the internal validation set. (K) ROC curve for patient survival of the external validation set. (L) Survival analysis between the two risk subgroups in the external validation set. GBM, glioblastoma; ROC, receiver operating characteristic; FABP5, fatty acid binding protein 5; ADRA1B, α-1B adrenergic receptor; AUC, area under the curve.

Journal: Molecular Medicine Reports

Article Title: Stigmasterol exerts antiglioma effects by regulating lipid metabolism

doi: 10.3892/mmr.2024.13351

Figure Lengend Snippet: Construction and validation of a prognostic model associated with stigmasterol target genes. (A) Venn diagram of potential targets of stigmasterol in the treatment of GBM, prognosis-related genes of patients with GBM and lipid metabolism-related genes. (B) Two prognosis-related target mRNAs of stigmasterol for GBM treatment were selected. (C and D) Least absolute shrinkage and selection operator variable screening process. (E) Distribution of survival status and risk score of patients with GBM in the training set. (F) Expression characteristics of the two prognosis-related target mRNAs of stigmasterol for GBM treatment in the training set. (G) ROC curve for patient survival of the training set. (H) Survival analysis between the two risk subgroups in the training set. (I) ROC curve for patient survival of the internal validation set. (J) Survival analysis between the two risk subgroups in the internal validation set. (K) ROC curve for patient survival of the external validation set. (L) Survival analysis between the two risk subgroups in the external validation set. GBM, glioblastoma; ROC, receiver operating characteristic; FABP5, fatty acid binding protein 5; ADRA1B, α-1B adrenergic receptor; AUC, area under the curve.

Article Snippet: For detection, the following secondary antibodies were used: HRP-conjugated goat anti-rabbit immunoglobulin G (IgG) antibody (dilution, 1:5,000; cat. no. A0208; Beyotime Institute of Biotechnology) for FABP5 and ADRA1B detection, and HRP-conjugated goat anti-mouse IgG antibody (dilution, 1:5,000, cat. no. SA00001-1; Proteintech Group, Inc.) for β-actin detection.

Techniques: Selection, Expressing, Binding Assay

Effects of stigmasterol on lipid metabolism and the expression of related target genes in GBM cells. (A) Effect of stigmasterol on free fatty acid levels in GBM cells. (B) Effect of stigmasterol on total cholesterol levels in GBM cells. (C) Effect of stigmasterol on the mRNA expression of the two target genes (FABP5 and ADRA1B) in GBM cells. (D) Effect of stigmasterol on the expression of the proteins encoded by the two target genes in GBM cells. (E) Molecular docking of the protein encoded by FABP5 with stigmasterol. (F) Molecular docking of the protein encoded by ADRA1B with stigmasterol. *P<0.05 and **P<0.01. GBM, glioblastoma; FABP5, fatty acid binding protein 5; ADRA1B, α-1B adrenergic receptor.

Journal: Molecular Medicine Reports

Article Title: Stigmasterol exerts antiglioma effects by regulating lipid metabolism

doi: 10.3892/mmr.2024.13351

Figure Lengend Snippet: Effects of stigmasterol on lipid metabolism and the expression of related target genes in GBM cells. (A) Effect of stigmasterol on free fatty acid levels in GBM cells. (B) Effect of stigmasterol on total cholesterol levels in GBM cells. (C) Effect of stigmasterol on the mRNA expression of the two target genes (FABP5 and ADRA1B) in GBM cells. (D) Effect of stigmasterol on the expression of the proteins encoded by the two target genes in GBM cells. (E) Molecular docking of the protein encoded by FABP5 with stigmasterol. (F) Molecular docking of the protein encoded by ADRA1B with stigmasterol. *P<0.05 and **P<0.01. GBM, glioblastoma; FABP5, fatty acid binding protein 5; ADRA1B, α-1B adrenergic receptor.

Article Snippet: For detection, the following secondary antibodies were used: HRP-conjugated goat anti-rabbit immunoglobulin G (IgG) antibody (dilution, 1:5,000; cat. no. A0208; Beyotime Institute of Biotechnology) for FABP5 and ADRA1B detection, and HRP-conjugated goat anti-mouse IgG antibody (dilution, 1:5,000, cat. no. SA00001-1; Proteintech Group, Inc.) for β-actin detection.

Techniques: Expressing, Binding Assay

The expression of specific transcription factors, membrane-associated proteins and cell markers in human small and large intestine. A Immunoblot of protein FOXM1, MYBL2 and UBE2C in villus and crypt derived from mouse small intestine. B FOXM1 and KI67 immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. C UBE2C immunostaining of large intestine derived from human specimens. Scale bars: 10 μm. D MYBL2 and KI67 immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. E Immunoblot of protein ANPEP, SLC12A2, IGF1R and SLC16A1 in villus and crypt derived from mouse small intestine. F SLC12A2 immunostaining of small intestine derived from human specimens. Scale bars: 50 μm. G SLC16A1 and E-Cadherin immunostaining of small intestine derived from human specimens. Scale bars: 50 μm. H ANPEP and E-Cadherin immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. I IGF1R immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. J Immunoblot of protein HSPD1, CPE, FABP5 and C1QBP in villus and crypt derived from mouse small intestine. K CPE and Chromogranin A immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. L HSPD1 and E-Cadherin immunostaining of small intestine derived from human specimens. Scale bars: 50 μm. M FABP5 and Chromogranin A immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. N C1QBP and E-Cadherin immunostaining of small intestine derived from human specimens. Scale bars: 50 μm

Journal: Cell Regeneration

Article Title: Identification of feature genes in intestinal epithelial cell types

doi: 10.1186/s13619-024-00208-8

Figure Lengend Snippet: The expression of specific transcription factors, membrane-associated proteins and cell markers in human small and large intestine. A Immunoblot of protein FOXM1, MYBL2 and UBE2C in villus and crypt derived from mouse small intestine. B FOXM1 and KI67 immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. C UBE2C immunostaining of large intestine derived from human specimens. Scale bars: 10 μm. D MYBL2 and KI67 immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. E Immunoblot of protein ANPEP, SLC12A2, IGF1R and SLC16A1 in villus and crypt derived from mouse small intestine. F SLC12A2 immunostaining of small intestine derived from human specimens. Scale bars: 50 μm. G SLC16A1 and E-Cadherin immunostaining of small intestine derived from human specimens. Scale bars: 50 μm. H ANPEP and E-Cadherin immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. I IGF1R immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. J Immunoblot of protein HSPD1, CPE, FABP5 and C1QBP in villus and crypt derived from mouse small intestine. K CPE and Chromogranin A immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. L HSPD1 and E-Cadherin immunostaining of small intestine derived from human specimens. Scale bars: 50 μm. M FABP5 and Chromogranin A immunostaining of small and large intestine derived from human specimens. Scale bars: 50 μm. N C1QBP and E-Cadherin immunostaining of small intestine derived from human specimens. Scale bars: 50 μm

Article Snippet: Mouse anti-Ki67 (1:200; 9449, CST), mouse anti-E-cadherin (1:1000; 610,182, BD Biosciences), mouse anti-chromogranin A (1:200; sc-393941, Santa Cruz), rabbit anti-Foxm1 (1:100; ab207298, Abcam), rabbit anti-Ube2c (1:50; 12,134–2-AP, Proteintech), rabbit anti-Mybl2 (1:100; ab76009, Abcam), rabbit anti-Anpep (1:50; 14,553–1-AP, Proteintech), rabbit anti-Slc12a2 (1:50; 13,884–1-AP, Proteintech), rabbit anti-Slc16a1 (1:50; 20,139–1-AP, Proteintech), rabbit anti-Igf1r (1:100; ab182408, Abcam), rabbit anti-Cpe (1:50; 13,710–1-AP, Proteintech), rabbit anti-Hspd1 (1:50; 15,282–1-AP, Proteintech), rabbit anti-C1qbp (1:50; 24,474–1-AP, Proteintech), rabbit anti-Fabp5 (1:100; ab255276, Abcam).

Techniques: Expressing, Membrane, Western Blot, Derivative Assay, Immunostaining