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

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

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

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

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

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

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

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

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

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

Journal: bioRxiv

Article Title: Lysosomes cell autonomously regulate myeloid cell states and immune responses

doi: 10.1101/2024.11.11.623074

Figure Lengend Snippet: Grn KO myeloid cells display an MG8-like signature in vitro . (a) Average expression of top 150 regulated genes [from Grn KO primary microglia (pMG) in vitro ] in each microglia subcluster from aged Grn KO microglia scRNA-seq. (b) Heatmap of relative expression of top 175 MG8 markers (from aged Grn KO microglia scRNA-seq) in WT and Grn KO pMG in vitro . (c) Volcano plot displaying differential expression between Grn KO and WT pMG. Top MG8 marker genes are highlighted as triangles. (d,e) Western blots and quantification of MG8 markers (GPNMB, FABP5, and total CatB) and LPL (not expressed by MG8) from protein lysates of WT and Grn KO mouse primary microglia (pMG) (n=5 biological replicates/genotype). (f) Average expression of top 150 upregulated genes [from Grn KO bone marrow-derived macrophages (BMDMs) in vitro ] in each microglia subcluster from aged Grn KO microglia scRNA-seq. (g) Heatmap of relative expression of top 175 MG8 markers (from aged Grn KO microglia scRNA-seq) in WT and Grn KO BMDMs in vitro . (h) Volcano plot displaying differential expression between Grn KO and WT BMDMs. Top MG8 marker genes are highlighted as triangles. (i) Western blots and quantification of MG8 markers (GPNMB, FABP5, and total CatB) and LPL (not expressed by MG8) from protein lysates of WT and Grn KO mouse BMDMs (n=3 biological replicates/genotype). Data presented in (e,j) are mean±s.e.m. Student’s t-tests were performed to compare across genotypes in (e,j). * P <0.05, ** P <0.01, *** P <0.001, **** P <0.0001.

Article Snippet: The following primary antibodies were used: sheep anti-PGRN (R&D Systems; Cat# 2557; 0.5μg/mL), rabbit anti-GPNMB (Cell Signaling Technology; Cat# 90205; 1:500), rabbit anti-FABP5 (Cell Signaling Technology; Cat# 39926; 1:500), rabbit anti-CatB (Cell Signaling Technology; Cat# 31718; 1:500), rabbit anti-LPL (Biotechne; Cat# AF7197; 1μg/mL), mouse anti-Vinculin (Sigma-Aldrich; Cat# V9264; 1:1000), mouse anti-β-Actin (Sigma-Aldrich; Cat# A2228; 1:10000), mouse anti-LC3 (Cell Signaling Technology; Cat# 83506; 1:500), mouse anti-p62 (Biotechne; Cat# MAB8028; 2μg/mL), mouse anti-AMPKα (Cell Signaling Technology; Cat# 2793; 1:500), rabbit anti-phospho-AMPKα (T172) (Cell Signaling Technology; Cat# 2535; 1:500), mouse anti- GAPDH (Abcam; Cat# ab8245; 1:2000), rabbit anti-ATP6V1A (Cell Signaling Technology; Cat# 39517; 1:500), rabbit anti-NPC1 (Abcam; Cat# ab134113; 1:500), rabbit anti-VPS34 (Cell Signaling Technology; Cat# 4263; 1:500), rabbit anti-CatD (Cell Signaling Technology; Cat# 88239; 1:500), sheep anti-TREM2 (Biotechne; Cat# AF1729; 0.1μg/mL), rabbit anti-ATG7 (Cell Signaling Technology; Cat# 8558; 1:500), rabbit anti-ATG14 (Cell Signaling Technology; Cat# 96752; 1:500), and rabbit anti-UVRAG (Abcam; Cat# ab313627; 1:500).

Techniques: In Vitro, Expressing, Marker, Western Blot, Derivative Assay

Journal: International Journal of Molecular Sciences

Article Title: Expression Profiles of Fatty Acid Transporters and the Role of n-3 and n-6 Polyunsaturated Fatty Acids in the Porcine Endometrium

doi: 10.3390/ijms252011102

Figure Lengend Snippet: Effect of n-6 (linoleic acid [LA] and arachidonic acid [ARA]; grey bars) and n-3 (α-linolenic acid [ALA], docosahexaenoic acid [DHA], and eicosapentaenoic acid [EPA]; black bars) PUFAs on the relative mRNA expression of genes involved in fatty acid binding ( FABP3 , FABP4 , FABP5 ; a ), action ( PPARA , PPARD , PPARG ; b ), and metabolism ( ACOX1 , CPT1A ; c ) as well as genes related to immune response ( IL1B , IL6 , TNF ; d ) and angiogenesis ( VEGFA , FGF2 ; e ) in the endometrium. Values from real-time PCR were normalized to geometric averaging of glyceraldehyde-3-phosphate dehydrogenase ( GAPDH ) and hypoxanthine phosphoribosyltransferase 1 ( HPRT1 ) mRNA expression. Data are expressed as means ± SEM ( n = 5). Asterisks specify the differences compared with the control value (CTRL; *, p ≤ 0.05; **, p < 0.01).

Article Snippet: To evaluate CD36 , SLC27A1 , SLC27A2 , SLC27A3 , SLC27A4 , SLC27A6 , PTGES , PTGIS , FABP3 , FABP4 , FABP5 , PPARA , PPARD , PPARG , ACOX1 , CPT1A , VEGFA , FGF2 , IL1B , IL6 , TNF , HPRT1 , GAPDH , and ACTG1 gene expression, 15 ng of complementary cDNA was amplified using TaqMan Gene Expression assays (Applied Biosystems by Thermo Fisher Scientific).

Techniques: Expressing, Binding Assay, Real-time Polymerase Chain Reaction, Control