Journal: Advanced Science

Article Title: HMGB2–RAD21 Axis Promotes Fibro/Adipogenic Progenitor Proliferation and Regulates Fat Infiltration

doi: 10.1002/advs.202514363

Figure Lengend Snippet: HMGB2 directly binds to the RAD21 promoter and activates its transcription. (A) Trends in HMGB2 and RAD21 expression in FAPs developmental trajectories. (B, C) EDU staining of mouse FAPs after transfection with si‐HMGB2. (D) The mRNA levels of HMGB2, RAD21, Cyclin E1, and Cyclin D1 were detected by qPCR ( n = 3) after transfection with si‐HMGB2. (E, F) EDU staining of mouse FAPs after transfection with si‐RAD21. (G) The mRNA levels of HMGB2, RAD21, Cyclin E1, and Cyclin D1 were detected by qPCR ( n = 3) after transfection with si‐RAD21. (H) EDU staining of mouse FAPs after transfection with si‐HMGB and pCDNA3.1‐RAD21. (I) Quantification of EDU + cells. (J) The mRNA levels of HMGB2, RAD21, Cyclin E1, and Cyclin D1 were detected by qPCR ( n = 3). (K) Real‐time proliferation index of FAPs. (L) Prediction of HMGB2 binding sites within the RAD21 promoter region (−2000 to +0bp) using the JASPAR database. (M) ChIP‐qPCR analyses of HMGB2 enrichment on RAD21 promoter in control and KLF4 knockdown cells. Data were normalized as a percentage of the input. (N) Experimental design involving amplification of the RAD21 promoter sequence from −2000 to +0 bp, deletion of the predicted binding region "ATATTA," and insertion into a promoter‐driven dual luciferase reporter plasmid pCIG vector carrying both firefly luciferase (Luc) and sea pansy luciferase (RLuc) (O) Dual luciferase reporter gene assay performed in 293T cells transfected with reporter plasmids containing wild‐type or mutant RAD21 promoters, along with pCDNA3.1‐HMGB2 vector ( n = 3).

Article Snippet: Real‐time quantitative PCR (qPCR) analyses were conducted on a LightCycler 480 II system (Roche, Basel, Switzerland) utilizing SYBR Green qPCR Mix (GDSBio, Guangzhou, China), with β‐actin serving as an internal control.

Techniques: Expressing, Staining, Transfection, Binding Assay, ChIP-qPCR, Control, Knockdown, Amplification, Sequencing, Luciferase, Plasmid Preparation, Reporter Gene Assay, Mutagenesis

Journal: Molecular Nutrition & Food Research

Article Title: Shielding Human Adipocytes From Inflammation: The Protective Potential of Polyphenol‐Rich Opuntia ficus‐indica Cladode Extract

doi: 10.1002/mnfr.70114

Figure Lengend Snippet: OFI‐CE treatment attenuates TNF‐α‐induced expression of IL‐6 and IL‐8. SGBSs were treated with OFI‐CE at the concentrations indicated for 4 h and then either treated with 10 ng/mL TNF‐α or left untreated for 18 h. Total RNA was extracted from cells, and mRNA levels of IL‐6 and IL‐8 were measured by qPCR using specific primers and normalized to 18S RNA. Data (means ± SD, n = 3) are expressed as percentage over TNF‐α alone. ## p < 0.01 versus basal (untreated) control; * p < 0.05 versus TNF‐α alone; ** p < 0.01 versus TNF‐α alone. For statistical analysis, we used Student's t test. IL‐6, interleukin‐6; IL‐8, interleukin‐8; OFI‐CE, Opuntia ficus‐indica (L.) Mill; TNF‐α, tumor necrosis factor‐α.

Article Snippet: Quantitative real‐time PCR (qPCR) analyses were performed with the CFX96 Touch Real‐Time PCR Detection System instrument and software (Bio‐Rad Laboratory).

Techniques: Expressing, Control

Journal: Molecular Nutrition & Food Research

Article Title: Shielding Human Adipocytes From Inflammation: The Protective Potential of Polyphenol‐Rich Opuntia ficus‐indica Cladode Extract

doi: 10.1002/mnfr.70114

Figure Lengend Snippet: OFI‐CE treatment counteracts TNF‐α‐induced expression of chemoattractant genes. SGBSs were treated with OFI‐CE at the concentrations indicated for 4 h and then either treated with 10 ng/mL TNF‐α or left untreated for 18 h. Total RNA was extracted from cells, and mRNA levels of MCP‐1, CXCL‐10, and M‐CSF were measured by qPCR using specific primers and normalized to 18S RNA. Data (means ± SD, n = 3) are expressed as percentage over TNF‐α alone. ## p < 0.01 versus basal (untreated) control; ** p < 0.01 versus TNF‐α alone. For statistical analysis, we used Student's t test. OFI‐CE, CXC‐L10, C‐X‐C motif chemokine ligand 10; MCP‐1, monocyte chemoattractant protein‐1; M‐CSF, macrophage colony‐stimulating factor; Opuntia ficus‐indica (L.) Mill; TNF‐α, tumor necrosis factor‐α.

Article Snippet: Quantitative real‐time PCR (qPCR) analyses were performed with the CFX96 Touch Real‐Time PCR Detection System instrument and software (Bio‐Rad Laboratory).

Techniques: Expressing, Control

Journal: Molecular Nutrition & Food Research

Article Title: Shielding Human Adipocytes From Inflammation: The Protective Potential of Polyphenol‐Rich Opuntia ficus‐indica Cladode Extract

doi: 10.1002/mnfr.70114

Figure Lengend Snippet: OFI‐CE counteracts TNF‐α‐induced ICAM‐1 expression and monocytes adhesion to inflamed adipocytes. SGBSs were treated with OFI‐CE at the concentrations indicated for 4 h and then either treated with 10 ng/mL TNF‐α or left untreated for 18 h. (A) Total RNA was extracted from cells, and mRNA levels of ICAM‐1 were measured by qPCR using specific primers and normalized to 18S RNA. Data (means ± SD, n = 3) are expressed as percentage over TNF‐α alone. ## p < 0.01 versus basal (untreated) control; * p < 0.05 versus TNF‐α alone; ** p < 0.01 versus TNF‐α alone. (B, C) Fluorescently labeled THP‐1 (106 cells/mL) was added to the SGBS monolayers. After 1 h, nonadhering cells were removed by three washes, and images of SGBS and adherent calcein‐labeled THP‐1 cells were visualized and captured with a fluorescent microscope at 10× magnification. (a) Control, (b) TNF‐α 10 ng/mL, (c) OFI‐CE 50 µg/mL + TNF‐α, (d) OFI‐CE 100 µg/mL + TNF‐α, and (e) OFI‐CE 200 µg/mL + TNF‐α. Data (means ± SD, n = 3) are expressed as the number of adherent monocytes over TNF‐α alone. ## p < 0.01 versus basal (untreated) control; * p < 0.05 versus TNF‐α alone. For statistical analysis, we used Student's t test. ICAM‐1, Intercellular adhesion molecule‐1; MTT, 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide; OFI‐CE, Opuntia ficus‐indica (L.) Mill; TNF‐α, tumor necrosis factor‐α.

Article Snippet: Quantitative real‐time PCR (qPCR) analyses were performed with the CFX96 Touch Real‐Time PCR Detection System instrument and software (Bio‐Rad Laboratory).

Techniques: Expressing, Control, Labeling, Microscopy