Journal: Frontiers in Endocrinology

Article Title: Dapagliflozin Attenuates Renal Tubulointerstitial Fibrosis Associated With Type 1 Diabetes by Regulating STAT1/TGFβ1 Signaling

doi: 10.3389/fendo.2019.00441

Figure Lengend Snippet: Sequences of primers used for quantitative RT-PCR analysis.

Article Snippet: Briefly, 4-μm-thick kidney sections were incubated with anti-FN (1:500), anti-Col IV (1:500), anti-STAT1 (1:500), and anti-TGFβ1 antibody (1:150) overnight at 4°C, followed by incubation with horseradish peroxidase-conjugated anti-rabbit secondary antibody (BBI Life Science, Shanghai, China).

Techniques: Quantitative RT-PCR

Journal: Frontiers in Endocrinology

Article Title: Dapagliflozin Attenuates Renal Tubulointerstitial Fibrosis Associated With Type 1 Diabetes by Regulating STAT1/TGFβ1 Signaling

doi: 10.3389/fendo.2019.00441

Figure Lengend Snippet: The expressions of STAT1, TGFβ1, FN, and Col IV in non-diabetic adjacent normal kidney tissues (NC), and renal biopsy sections from DN patients. (A) Protein expression levels of STAT1, TGFβ1, FN, and Col IV were quantified by western blot analysis. (B) mRNA expressions levels of STAT1, TGFβ1, FN, and Col IV were quantified by qRT-PCR. * P < 0.05 vs. NC.

Article Snippet: Briefly, 4-μm-thick kidney sections were incubated with anti-FN (1:500), anti-Col IV (1:500), anti-STAT1 (1:500), and anti-TGFβ1 antibody (1:150) overnight at 4°C, followed by incubation with horseradish peroxidase-conjugated anti-rabbit secondary antibody (BBI Life Science, Shanghai, China).

Techniques: Expressing, Western Blot, Quantitative RT-PCR

Journal: Frontiers in Endocrinology

Article Title: Dapagliflozin Attenuates Renal Tubulointerstitial Fibrosis Associated With Type 1 Diabetes by Regulating STAT1/TGFβ1 Signaling

doi: 10.3389/fendo.2019.00441

Figure Lengend Snippet: Effect of DAPA on renal production of fibrotic markers in diabetic mice. (A) Representative western blots for STAT1, TGF-β1, Col IV, and FN in renal cortical lysates. β-actin was used as internal control. (B) Density ratio of STAT1 to GAPDH mRNA expressions. (C–F) Normalized quantification of western blot data in diabetic mice at different times. * P < 0.05 vs. NC; ** P < 0.05 vs. DM-8W; * & P < 0.05 vs. DM-12W; # P < 0.05 vs. DM-16W. Values are means ± SEM. n = 3.

Article Snippet: Briefly, 4-μm-thick kidney sections were incubated with anti-FN (1:500), anti-Col IV (1:500), anti-STAT1 (1:500), and anti-TGFβ1 antibody (1:150) overnight at 4°C, followed by incubation with horseradish peroxidase-conjugated anti-rabbit secondary antibody (BBI Life Science, Shanghai, China).

Techniques: Western Blot, Control

Journal: Frontiers in Endocrinology

Article Title: Dapagliflozin Attenuates Renal Tubulointerstitial Fibrosis Associated With Type 1 Diabetes by Regulating STAT1/TGFβ1 Signaling

doi: 10.3389/fendo.2019.00441

Figure Lengend Snippet: Effects of DAPA and Flu on the expression of EMT-related proteins in HK-2 cells. (A) Representative western blots illustrating STAT1, TGF-β1, E-cadherin, α-SMA, and β-actin protein expression in HK2 cells. (B–E) Quantification of the western blot data in HK-2 cells. Statistical analysis was performed with one-way ANOVA. Results are expressed as the mean ± SEM. * P < 0.05 vs. NC, # P < 0.05 vs. HG, #& P < 0.05 vs. HG+DAPA-1.

Article Snippet: Briefly, 4-μm-thick kidney sections were incubated with anti-FN (1:500), anti-Col IV (1:500), anti-STAT1 (1:500), and anti-TGFβ1 antibody (1:150) overnight at 4°C, followed by incubation with horseradish peroxidase-conjugated anti-rabbit secondary antibody (BBI Life Science, Shanghai, China).

Techniques: Expressing, Western Blot

Journal: Frontiers in Immunology

Article Title: Oxygen Desaturation Is Associated With Fibrocyte Activation via Epidermal Growth Factor Receptor/Hypoxia-Inducible Factor-1α Axis in Chronic Obstructive Pulmonary Disease

doi: 10.3389/fimmu.2022.852713

Figure Lengend Snippet: Higher percentage of circulating fibrocytes with greater lung function decline in COPD desaturators (A) . NANT cells were isolated from peripheral blood of non-desaturators (n = 12) and desaturators (n = 12) and the percentage of collagen I + /CD45 + fibrocyte in freshly isolated NANT cells on day 0 was determined by flow cytometry. Horizontal lines represent the mean values for each group. (B–F) All participants performed 6-minute walking tests at the beginning and during the 5-year follow-up. The correlation between the percentage of fibrocytes and the extent of post-exercise reduction in SpO 2 at baseline (B) , annualized change in FEV 1 /FVC ratio (C) , FEV 1 (D) , FVC (E) and 6MWD (F) are shown. The differences between non-desaturators and desaturators were determined by Mann-Whitney test and the correlation was determined by Spearman’s rank correlation. **** P < 0.0001. COPD, chronic obstructive pulmonary disease; NANT cells, non-adherent non-T fraction of peripheral blood mononuclear cells; FEV 1 , forced expiratory volume in 1 second; FVC, forced vital capacity; %pred, percent of predicted value; Ex, exercise; SpO 2 , peripheral blood oxygen saturation; 6MWD, 6-minute walking distance.

Article Snippet: Permeabilised NANT cells were incubated with FITC-conjugated rabbit anti-human CTGF antibodies (Abcam, Cambridge, UK), followed by staining with anti-COL I-PE (Merck Milli-Mark, CA, USA) and anti-CD45-PerCp polyclonal antibodies to evaluate the expression of CTGF in the fibrocytes.

Techniques: Isolation, Flow Cytometry, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Oxygen Desaturation Is Associated With Fibrocyte Activation via Epidermal Growth Factor Receptor/Hypoxia-Inducible Factor-1α Axis in Chronic Obstructive Pulmonary Disease

doi: 10.3389/fimmu.2022.852713

Figure Lengend Snippet: Identification of COL I + /CD34 + fibrocytes in the lung tissue of COPD desaturators (A–C) . The paraffin sections of lung tissue from COPD desaturators were stained for the CD34 surface marker ( green ), intracellular COL I ( red ), and nuclei ( blue ) (A) , and for hematoxylin and eosin staining (B) . The immunofluorescence staining and confocal microscopic analysis are described in Methods. (C) Shows a magnification of the merged image of (A) , and arrows indicate the double staining of COL I + /CD34 + fibrocytes with yellow enhancement in the merged image. Bar , 75 μm (A) and 25μm (C) . (D, E) The paraffin sections of airway tissue from COPD non-desaturators were stained for indicated immunofluorescence staining (D) and hematoxylin and eosin staining (E) , as described above. Bar , 75 μm (D) and 30 μm (E) . (F) The differences between the numbers of fibrocytes in specimens from non-desaturators and desaturators were determined by Mann-Whitney test. ** P < 0.01. COL I, collagen I; COPD, chronic obstructive pulmonary disease; In, interstitium; Ep, epithelium; En, endothelium; Al, alveoli.

Article Snippet: Permeabilised NANT cells were incubated with FITC-conjugated rabbit anti-human CTGF antibodies (Abcam, Cambridge, UK), followed by staining with anti-COL I-PE (Merck Milli-Mark, CA, USA) and anti-CD45-PerCp polyclonal antibodies to evaluate the expression of CTGF in the fibrocytes.

Techniques: Staining, Marker, Immunofluorescence, Double Staining, MANN-WHITNEY

Journal: Frontiers in Immunology

Article Title: Oxygen Desaturation Is Associated With Fibrocyte Activation via Epidermal Growth Factor Receptor/Hypoxia-Inducible Factor-1α Axis in Chronic Obstructive Pulmonary Disease

doi: 10.3389/fimmu.2022.852713

Figure Lengend Snippet: Higher CXCR4, CTGF, EGFR and HIF-1α expression with greater increased in number and myofibroblastic differentiation in fibrocytes from COPD desaturators. The number of Col I + /CD45 + fibrocytes (A) and differentiating α-SMA + fibrocytes (B) in NANT cells were determined by flow cytometry after 14 days in culture. NANT cells were isolated from COPD non-desaturators and desaturators. The proportion of CXCR4- (C) , CTGF- (D) , EGFR- (E) and HIF-1α- (F) expressing fibrocytes within freshly isolated NANT cells from desaturators, determined by flow cytometry, was higher compared to that from non-desaturators. Horizontal lines represent the median values for each group. The differences between disease groups were determined by Mann-Whitney test. * P < 0.05, ** P < 0.01, *** P < 0.001. CXCR4, CXC chemokine receptor 4; CTGF, connective tissue growth factor; EGFR, epidermal growth factor receptor; HIF-1α, hypoxia-inducible factor-1α; COPD, chronic obstructive pulmonary disease; NANT cells, non-adherent non-T fraction of peripheral blood mononuclear cells; α-SMA, α-smooth muscle actin.

Article Snippet: Permeabilised NANT cells were incubated with FITC-conjugated rabbit anti-human CTGF antibodies (Abcam, Cambridge, UK), followed by staining with anti-COL I-PE (Merck Milli-Mark, CA, USA) and anti-CD45-PerCp polyclonal antibodies to evaluate the expression of CTGF in the fibrocytes.

Techniques: Expressing, Flow Cytometry, Isolation, MANN-WHITNEY