Journal: Gut Microbes

Article Title: Microbial metabolite Enterobactin impairs mitochondrial respiration and alleviates colitis

doi: 10.1080/19490976.2026.2662638

Figure Lengend Snippet: Monomeric Ent, 2, 3-DHBA, but not 2, 5-DHBA, restores mucosal integrity and mitochondrial homeostasis in DSS-induced colitis. Acute colitis was induced in mice as in Figure 7. After the cessation of DSS (recovery phase) mice were given either 2, 3-DHBA or 2, 5-DHBA (500 µg/mouse per day) for 7 d and euthanized. (A) Representative H&E-stained colon sections showing epithelial damage, crypt loss, and inflammatory cells ( n = 5/group). (B) Alcian blue staining for goblet cells in the colon (C) Immunoblot analysis of tight junction proteins (occludin and claudin-1) in colonic tissue. (D) qRT-PCR analysis of genes involved in mitochondrial biogenesis peroxisome proliferator activated receptor gamma coactivator 1-alpha ( PGC-1α ) and deiodinase type 2 ( Dio2) .(E) Colonic expression of antioxidant genes (Superoxide dismutase 2 (SOD2) , glutathione peroxidase (Gpx) , and nuclear factor erythroid 2-related factor 2 (Nrf2) (F) Immunoblot showing protein involved in mitochondrial dynamics uncoupling protein 1 (Ucp1), mitofusin 1 (Mfn1), and optic atrophy type 1 (Opa1). Data is presented as mean ± SEM ( n = 5). Statistical comparisons were performed using one-way ANOVA with Tukey’s post hoc test (* p < 0.05, ** p < 0.01, and *** p < 0.001).

Article Snippet: Antibodies to OXPHOS (Catalog # 45-8099) were from Thermo Fischer, and antibodies to occludin (Catalog #40-4700) and claudin-1 (Catalog #37-4900) from Invitrogen and UCP1 (Catalog #MAB6158) from R&D Systems.

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

Journal: Cells

Article Title: Toward Efficient Beige Adipogenesis: Protocol Optimization Using Adipose-Derived Stem Cells

doi: 10.3390/cells15010054

Figure Lengend Snippet: Comparative analysis of changes in relative UCP1 gene expression at the mRNA level, normalized to the average GAPDH gene expression, depending on the duration of the differentiation-inducing culture of ADSCs. ADSCs were cultured for 4 days (IND-4D DEX), 8 days (IND-8D DEX), or 21 days (IND-21D) in induction medium supplemented with 10 μg/mL insulin, 0.2 μM rosiglitazone, 1 μM dexamethasone, and 500 μM IBMX followed by culture without IBMX until day 21 post-T0. Fold change was calculated relative to control using the 2^−ΔΔCT method. Statistical analysis was performed using ANOVA followed by Tukey’s post hoc test, *** p < 0.001 compared to the control, # p < 0.05 compared to IND-8D DEX.

Article Snippet: The membrane was incubated overnight at 4 °C with mouse anti-UCP1 antibody (R&D Systems, Minneapolis, MI, USA) at a concentration of 0.5 μg/mL and rabbit anti-GAPDH antibody (Merck KGaA, Darmstadt, Germany) at a concentration of 0.1 μg/mL.

Techniques: Gene Expression, Cell Culture, Control

Journal: Cells

Article Title: Toward Efficient Beige Adipogenesis: Protocol Optimization Using Adipose-Derived Stem Cells

doi: 10.3390/cells15010054

Figure Lengend Snippet: UCP1 gene expression at the mRNA level, normalized to the average GAPDH gene expression in ADSCs exposed to varying concentrations of rosiglitazone, compared with control cells. ADSCs were differentiated for 8 days in induction medium supplemented with 10 μg/mL insulin, 1 μM dexamethasone, 500 μM IBMX, and rosiglitazone at the following concentrations: 0.2 μM (ROSI-0.2), 1 μM (ROSI-1), 5 μM (ROSI-5), and 10 μM (ROSI-10) followed by culture without IBMX until day 21 post-T0. Fold change was calculated relative to control using the 2^−ΔΔCT method. Statistical analysis was performed using ANOVA followed by Tukey’s post hoc test; * p < 0.05, *** p < 0.001 compared to control, ### p < 0.001 compared to ROSI-1, ++ p < 0.005, +++ p < 0.001 compared to ROSI-5, ^^^ p < 0.001 compared to ROSI-10.

Article Snippet: The membrane was incubated overnight at 4 °C with mouse anti-UCP1 antibody (R&D Systems, Minneapolis, MI, USA) at a concentration of 0.5 μg/mL and rabbit anti-GAPDH antibody (Merck KGaA, Darmstadt, Germany) at a concentration of 0.1 μg/mL.

Techniques: Gene Expression, Control

Journal: Cells

Article Title: Toward Efficient Beige Adipogenesis: Protocol Optimization Using Adipose-Derived Stem Cells

doi: 10.3390/cells15010054

Figure Lengend Snippet: Semi-quantitative analysis of ( A ) GAPDH and ( B ) UCP1 by Western blot in the ADSCs after differentiation towards brown adipose tissue using various protocols. WM- weight marker; ROSI-5, ROSI-1, ROSI-0.2—different concentrations of rosiglitazone (µM); IND-4D/IND-8D—4 or 8 days of induction culture; DR5 –maintenance medium with rosiglitazone and dexamethasone; R5—maintenance medium with rosiglitazone, INS-5, INS-10, INS-20—different concentrations of insulin (μg/mL); 10% FBS, 5% FBS—different concentration of fetal bovine serum in adipogenic medium; CONTROL/CONTROL 21D—ADSCs not subjected to the differentiation process collected immediately after reaching confluency/cultured for three weeks post-confluency.

Article Snippet: The membrane was incubated overnight at 4 °C with mouse anti-UCP1 antibody (R&D Systems, Minneapolis, MI, USA) at a concentration of 0.5 μg/mL and rabbit anti-GAPDH antibody (Merck KGaA, Darmstadt, Germany) at a concentration of 0.1 μg/mL.

Techniques: Western Blot, Marker, Concentration Assay, Control, Cell Culture

Journal: Cells

Article Title: Toward Efficient Beige Adipogenesis: Protocol Optimization Using Adipose-Derived Stem Cells

doi: 10.3390/cells15010054

Figure Lengend Snippet: Comparative analysis of changes in relative UCP1 gene expression at the mRNA level, normalized to the average GAPDH gene expression, in primary ADSCs obtained from different adults and subjected to differentiation using the optimized protocol, compared to undifferentiated control cells. ADSCs were differentiated for 8 days in induction medium supplemented with 20 μg/mL insulin, 1 μM dexamethasone, 500 μM IBMX, and 5 μM rosiglitazone, followed by culture without IBMX until day 21 post-T0. Fold change was calculated relative to control using the 2^−ΔΔCT method. Statistical analysis was performed using ANOVA followed by Tukey’s post hoc test, * p < 0.05.

Article Snippet: The membrane was incubated overnight at 4 °C with mouse anti-UCP1 antibody (R&D Systems, Minneapolis, MI, USA) at a concentration of 0.5 μg/mL and rabbit anti-GAPDH antibody (Merck KGaA, Darmstadt, Germany) at a concentration of 0.1 μg/mL.

Techniques: Gene Expression, Control

Journal: Cells

Article Title: Toward Efficient Beige Adipogenesis: Protocol Optimization Using Adipose-Derived Stem Cells

doi: 10.3390/cells15010054

Figure Lengend Snippet: Semi-quantitative analysis of GAPDH ( A ) and UCP1 ( B ) by Western blot in the primary ADSCs collected from different donors, after differentiation towards brown adipose tissue using optimized protocol. WM- molecular weight marker; P7,P15,P58,P12—primary human ADSCs from different donors; AD—after differentiation, BD—before differentiation; PC—positive control.

Article Snippet: The membrane was incubated overnight at 4 °C with mouse anti-UCP1 antibody (R&D Systems, Minneapolis, MI, USA) at a concentration of 0.5 μg/mL and rabbit anti-GAPDH antibody (Merck KGaA, Darmstadt, Germany) at a concentration of 0.1 μg/mL.

Techniques: Western Blot, Molecular Weight, Marker, Positive Control