Journal: iScience

Article Title: Chidamide suppresses adipogenic differentiation of bone marrow derived mesenchymal stem cells via increasing REEP2 expression

doi: 10.1016/j.isci.2023.106221

Figure Lengend Snippet: CHI suppresses adipogenic differentiation of BM-MSCs in a dose-dependent manner (A) Chemical structure of CHI. (B and C) BM-MSCs were cultured with or without adipogenic medium for 8 days, and the differentiation induced BM-MSCs were also treated with different concentrations of CHI (0, 0.5, 1, 2, 3 μM). (B) The mRNA of adipogenic markers (CEBPA, FABP4, PLIN1, PPARG and LPL) were detected using qRT-PCR. GAPDH was used as a loading control. Data were represented as mean ± SEM. (C) Oil red O staining. The pictures were captured using Olympus microscope under 10× and 20× objective lenses and a representative experiment was presented. The AD group was used to compare with other groups. Scale bars, 100 μm/200 μm. ∗∗∗p<0.001 by Student’s t test. UN, untreated; AD, adipogenic differentiation; CHI, Chidamide.

Article Snippet: Human Bone Marrow Mesenchymal Stem Cells Osteogenic Differentiation Kit , Cyagen , Cat#HUXMX-90021.

Techniques: Cell Culture, Quantitative RT-PCR, Control, Staining, Microscopy

Journal: iScience

Article Title: Chidamide suppresses adipogenic differentiation of bone marrow derived mesenchymal stem cells via increasing REEP2 expression

doi: 10.1016/j.isci.2023.106221

Figure Lengend Snippet: Gene expression profiling of BM-MSCs toward adipogenic differentiation with or without CHI (A) The normalized intensity values of gene expression in the four BM-MSCs groups (UN, AD, DMSO, and CHI). (B and C) The distributions of gene expression in the four groups based on normalized values. (B) AD versus UN. (C) CHI versus DMSO. (D) The cluster characteristics in gene expression pattern of the four groups plotted by heatmap diagram. (E and F) The combined analysis of differentially expressed genes in adipogenic differentiation of BM-MSCs and CHI management. (E) Genes upregulated in AD versus UN and downregulated in CHI versus DMSO. (F) Genes downregulated in AD versus UN and upregulated in CHI versus DMSO. (G) The cluster analysis of the combined differential genes by heatmap diagram. (H and I) The gene ontology (GO) (H) and pathway (I) analysis of the combined differential genes.

Article Snippet: Human Bone Marrow Mesenchymal Stem Cells Osteogenic Differentiation Kit , Cyagen , Cat#HUXMX-90021.

Techniques: Gene Expression

Journal: iScience

Article Title: Chidamide suppresses adipogenic differentiation of bone marrow derived mesenchymal stem cells via increasing REEP2 expression

doi: 10.1016/j.isci.2023.106221

Figure Lengend Snippet: REEP2 was verified as a downstream target of CHI during adipogenic differentiation of BM-MSCs (A) The top 50 genes downregulated in AD and upregulated by CHI were shown. (B) qRT-PCR analysis of the five selected genes during adipogenic differentiation of BM-MSCs with or without CHI. Data were represented as mean ± SEM. (C) The REEP2 protein expression in adipogenic differentiation of BM-MSCs with or without CHI. (D) Ultrasonication efficiency of the genomic DNA detected by agarose gel electrophoresis. (E) Sketch map showing the distribution of primers used to expand the putative H3K27ac and H3K9ac binding sites in REEP2 promoter region. (F) ChIP-qRT-PCR analysis of H3K27ac and H3K9ac binding level in the REEP2 promoter region after CHI treatment. Data were represented as mean ± SEM. ∗p<0.05, ∗∗p<0.01, ∗∗∗p<0.001 by Student’s t test.

Article Snippet: Human Bone Marrow Mesenchymal Stem Cells Osteogenic Differentiation Kit , Cyagen , Cat#HUXMX-90021.

Techniques: Quantitative RT-PCR, Expressing, Agarose Gel Electrophoresis, Binding Assay

Journal: iScience

Article Title: Chidamide suppresses adipogenic differentiation of bone marrow derived mesenchymal stem cells via increasing REEP2 expression

doi: 10.1016/j.isci.2023.106221

Figure Lengend Snippet: REEP2 acts as a negative regulator of adipogenic differentiation of BM-MSCs (A) The mRNA and protein expression of REEP2 were detected in BM-MSCs infected with lentiviruses encoding GFP or shREEP2, followed by adipogenic induction for 8 days. (B) qRT-PCR and western blot analyses of REEP2 expression in BM-MSCs infected with lentiviruses expressing vector (A1) or REEP2 (A1-REEP2), followed by adipogenic induction for 8 days. (C and D) Expression analyses of adipogenic markers (CEBPA, FABP4, PLIN1, PPARG and LPL) were performed using qRT-PCR in adipogenic medium induced BM-MSCs after knock-down (C) and over-expression of REEP2 (D). (E) Oil red O staining of lentiviruses infected and adipogenic medium induced BM-MSCs in C and D. The pictures were captured using Olympus microscope under 10× and 20× objective lenses and a representative experiment was presented. Scale bars, 100 μm/200 μm. Data were represented as mean ± SEM. ∗p<0.05, ∗∗p<0.01, ∗∗∗p<0.001 by Student’s t test.

Article Snippet: Human Bone Marrow Mesenchymal Stem Cells Osteogenic Differentiation Kit , Cyagen , Cat#HUXMX-90021.

Techniques: Expressing, Infection, Quantitative RT-PCR, Western Blot, Plasmid Preparation, Knockdown, Over Expression, Staining, Microscopy

Journal: iScience

Article Title: Chidamide suppresses adipogenic differentiation of bone marrow derived mesenchymal stem cells via increasing REEP2 expression

doi: 10.1016/j.isci.2023.106221

Figure Lengend Snippet: REEP2 mediates the suppressive effect of CHI on adipogenic differentiation of BM-MSCs (A and B) The mRNA and protein expression of REEP2 were evaluated by qRT-PCR (A) and western blot (B), respectively, in BM-MSCs infected with lenti-shREEP2 (or lenti-GFP) followed by adipogenic induction with CHI treatment or DMSO. (C–G) Adipogenic markers CEBPA (C), FABP4 (D), PLIN1 (E), PPARG (F) and LPL (G) expression were detected in lentivirues infected BM-MSCs followed by adipogenic induction and simultaneous treatment with CHI or DMSO for 8 days. (H) Oil red O staining. The pictures were captured using Olympus microscope under 10× and 20× objective lenses and a representative experiment was presented. Scale bars, 100 μm/200 μm. Data were represented as mean ± SEM. ∗p< 0.05, ∗∗p< 0.01 and ∗∗∗p<0.001 by Student’s t test.

Article Snippet: Human Bone Marrow Mesenchymal Stem Cells Osteogenic Differentiation Kit , Cyagen , Cat#HUXMX-90021.

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Infection, Staining, Microscopy

Journal: iScience

Article Title: Chidamide suppresses adipogenic differentiation of bone marrow derived mesenchymal stem cells via increasing REEP2 expression

doi: 10.1016/j.isci.2023.106221

Figure Lengend Snippet:

Article Snippet: Human Bone Marrow Mesenchymal Stem Cells Osteogenic Differentiation Kit , Cyagen , Cat#HUXMX-90021.

Techniques: Recombinant, SYBR Green Assay, Protease Inhibitor, Stripping Membranes, cDNA Synthesis, Chromatin Immunoprecipitation, Bicinchoninic Acid Protein Assay, Plasmid Preparation, Purification, Expressing, Software