Journal: Journal of the American Society of Nephrology : JASN

Article Title: Specific Macrophage Subtypes Influence the Progression of Rhabdomyolysis-Induced Kidney Injury

doi: 10.1681/ASN.2014040320

Figure Lengend Snippet: The kidney macrophage subtype evolution mirrors kidney function. (A) The elevated BUN levels observed at day 2 (Figure 1F) decreased by day 8, indicating ongoing kidney repair (n=6–11). (B–D) Analysis of macrophages obtained from kidney cell suspensions. Three regions were discriminated according to the expression levels of CD11b and F4/80 as follows: F4/80-CD11b+ (R0), F4/80lowCD11bhigh (R1), and F4/80highCD11b+ (R2). CD11b corresponds to R0+R1+R2 sum. Macrophage distribution among the three regions was affected by glycerol injection at day 2 and day 8. (B) Representative dotplot gated on 30,000 live CD45+ cells. (C) R0, R1, R2, and total CD11b+ cell counts in kidney samples (n=3–5; **P<0.01 and ***P<0.001 compared with day 0; #P<0.05 and ####P<0.0001 compared with day 2). (D) Distribution of CD11b+ cells in the three regions (n=3–5). Macrophage subpopulations were characterized by expression of surface markers using flow cytometry. Mean fluorescence intensity (MFI) is reported on the yaxis for the following markers: (E) CD206 (M2 marker) and (F) CD36 (M2 marker). (n=3–9; *A star above a line displays P<0.05; #displays significant difference between R2 at day 2 or day 8 compared with R2 at day 0 [p<0.05]; $displays significant difference between R1 at day 8 compared with R1 at day 2 [p<0.05]. Lines display means±SEM for individual data).

Article Snippet: Specific primary antibodies were incubated on mouse tissue sections for the detection of F4/80 (rat anti-mouse, 1/100, clone BM8, MF48000; Invitrogen, Saint Aubin, France), collagen III (rabbit anti-mouse, BP8014; Acris Antibodies GmbH, Herford, Germany) or fibronectin (rabbit anti-human/mouse, F3648; Sigma-Aldrich, Saint-Quentin Fallavier, France).

Techniques: Expressing, Injection, Flow Cytometry, Fluorescence, Marker

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Specific Macrophage Subtypes Influence the Progression of Rhabdomyolysis-Induced Kidney Injury

doi: 10.1681/ASN.2014040320

Figure Lengend Snippet: CL-mediated macrophage depletion attenuates kidney lesions 2 days after glycerol injection. Mice received EL or lCL in the saline (NaCl) or the glycerol (Gly) condition according to the protocol shown in Figure 6E. (A and B) CL reduced glycerol-induced fibronectin (Original magnification, ×400 in A) and collagen III (Original magnification, ×200 in B) accumulation at the protein level (left panel; n=3–9) and at the mRNA level in the whole kidney (right panel; n=5–6). (C) Hmox1 was upregulated by glycerol treatment and not modified by CL. CL reduced glycerol-induced Ccl2 and Ccl7 mRNA expression (n=5–6). (Versus NaCl EL: ***P<0.001 and ****P<0.0001. Versus glycerol EL: ##P<0.01; ###P<0.001; ####P<0.0001.) (D) CL reduced glycerol-induced lesions on periodic acid-Schiff staining (Original magnification, ×200 in representative examples). (E) CL did not affect rhabdomyolysis intensity (n=4–8).

Article Snippet: Specific primary antibodies were incubated on mouse tissue sections for the detection of F4/80 (rat anti-mouse, 1/100, clone BM8, MF48000; Invitrogen, Saint Aubin, France), collagen III (rabbit anti-mouse, BP8014; Acris Antibodies GmbH, Herford, Germany) or fibronectin (rabbit anti-human/mouse, F3648; Sigma-Aldrich, Saint-Quentin Fallavier, France).

Techniques: Injection, Saline, Modification, Expressing, Staining

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Specific Macrophage Subtypes Influence the Progression of Rhabdomyolysis-Induced Kidney Injury

doi: 10.1681/ASN.2014040320

Figure Lengend Snippet: CL-mediated macrophage depletion attenuates kidney fibrosis. (A–D) One month after rhabdomyolysis. CTL, age-matched control group; Gly, glycerol-treated group; Gly CL, CL-treated mice according to the pre, post, and glycerol CL protocol depicted in Figure ​Figure6C.6C. (A) BUN levels (n=2–6). (B) CD206+ expression was significantly increased in the glycerol CL condition among CD11b+ cells (R0, R1, and R2) (n=5–8). (C) Collagen III and Masson trichrome staining (representative examples are shown). (D) Collagen III deposit quantification (n=5–7). (Versus control: **P<0.01 and ****P<0.0001. Versus Gly: #P<0.05 and ##P<0.01.) (E–H) Seven months after rhabdomyolysis. NaCl EL, glycerol EL, and pre-glycerol early CL refer to mice treated according to the protocol depicted in Figure 6A. (E) BUN levels (n=6–14). (F) Kidney mass indicated global atrophy after glycerol injection. (n=6–14) (G) Collagen III and Masson trichrome staining (representative examples are shown). (H) Collagen III deposits were still significantly increased in the glycerol condition, which was partially prevented by CL (n=6–14). (Versus NaCl EL: *P<0.05 and ****P<0.0001. Versus glycerol: #P<0.05, ##P<0.01, ####P<0.0001.)

Article Snippet: Specific primary antibodies were incubated on mouse tissue sections for the detection of F4/80 (rat anti-mouse, 1/100, clone BM8, MF48000; Invitrogen, Saint Aubin, France), collagen III (rabbit anti-mouse, BP8014; Acris Antibodies GmbH, Herford, Germany) or fibronectin (rabbit anti-human/mouse, F3648; Sigma-Aldrich, Saint-Quentin Fallavier, France).

Techniques: Control, Expressing, Staining, Injection

Journal: Journal of the American Society of Nephrology : JASN

Article Title: Specific Macrophage Subtypes Influence the Progression of Rhabdomyolysis-Induced Kidney Injury

doi: 10.1681/ASN.2014040320

Figure Lengend Snippet: Proposed early mechanisms involved in rhabdomyolysis-induced kidney injury. (1) In response to myoglobin, tubular cells secrete macrophage chemoattractants (Ccl2, Ccl7). (2) In response to these chemoattractants blood monocytes migrate to the renal interstitium. (3) In addition to its effect on tubular cells, myoglobin polarizes these macrophages toward a proinflammatory phenotype. (4) CD11bhighF4/80lowLy6-Bhigh subtype macrophages enhance renal injury by secreting extracellular matrix compounds (fibronectin, collagen III), proinflammatory cytokines (Il1b, Il12p40) and by increasing recruitment of newly generated macrophages (secretion of Ccl2 and Ccl7).

Article Snippet: Specific primary antibodies were incubated on mouse tissue sections for the detection of F4/80 (rat anti-mouse, 1/100, clone BM8, MF48000; Invitrogen, Saint Aubin, France), collagen III (rabbit anti-mouse, BP8014; Acris Antibodies GmbH, Herford, Germany) or fibronectin (rabbit anti-human/mouse, F3648; Sigma-Aldrich, Saint-Quentin Fallavier, France).

Techniques: Generated

Journal: Frontiers in Pharmacology

Article Title: Ivabradine Ameliorates Cardiac Diastolic Dysfunction in Diabetic Mice Independent of Heart Rate Reduction

doi: 10.3389/fphar.2021.696635

Figure Lengend Snippet: JNK and p38 MAPK mediate high glucose (HG)-induced fibrogenic protein upregulation in vitro (A) NRCFs were confirmed by vimentin and DAPI staining (original magnification × 400). Scale bars, 50 μm. Data were obtained from three independent experiments (B and C) NRCFs transfected with or without JNK or p38 MAPK siRNA were evaluated for the protein expression of JNK and p38 MAPK by western blotting (D) NRCFs were exposed to HG for 0–48 h and JNK/p38 MAPK phosphorylation and total JNK/p38 MAPK expression were determined by western blotting (E) NRCFs were transfected with or without JNK/p38 MAPK siRNA and then exposed to HG for the indicated periods of time. The protein expression of PCNA, α-SMA, TIMP2, collagen I, collagen III, and MMP2 was determined by western blotting. Data in (B–E) are presented as mean ± SEM ( n = 3). p -values in (B–D) were calculated using paired Student’s t -test. p -value in (E) was calculated using one-way ANOVA with Tukey multiple comparison test. * p < 0.05, compared to the normal group; # p < 0.05, compared to the HG group.

Article Snippet: Antibodies against collagen I and collagen III were obtained from the Biorbyt Corporation (Orwell Furlong, Cambridge, United Kingdom).

Techniques: In Vitro, Staining, Transfection, Expressing, Western Blot, Phospho-proteomics, Comparison

Journal: Frontiers in Pharmacology

Article Title: Ivabradine Ameliorates Cardiac Diastolic Dysfunction in Diabetic Mice Independent of Heart Rate Reduction

doi: 10.3389/fphar.2021.696635

Figure Lengend Snippet: JNK and p38 MAPK mediate the diabetes-induced increase in fibrogenic protein expression in vivo (A) Left ventricular fibroblasts isolated from mice were confirmed by vimentin staining. Data was obtained from three independent experiments (B and C) Isolated left ventricular fibroblasts from wild-type mice that were or were not injected with lentivirus via a tail vein were evaluated for JNK and p38 MAPK protein expression after 4 weeks (D) Left ventricular fibroblasts were isolated from mice and JNK and p38 MAPK phosphorylation and total JNK and p38 MAPK expression were determined by western blotting (E) Isolated left ventricular fibroblasts from diabetic mice that had or had not been injected with lentivirus via a tail vein were evaluated for PCNA, α-SMA, TIMP2, collagen I, collagen III, and MMP2 expression after 4 weeks (F) Between 7 and 11 weeks following the tail vein injection of lentivirus, the blood glucose concentration and body mass of the mice were measured. Data in (B–F) are presented as mean ± SEM ( n = 6). p -values in (B–D) were calculated using paired Student’s t -test. p -values in (E-F) were calculated using one-way ANOVA with Tukey multiple comparison test. * p < 0.05, compared to the Control group; # p < 0.05, compared to the db/db mice group.

Article Snippet: Antibodies against collagen I and collagen III were obtained from the Biorbyt Corporation (Orwell Furlong, Cambridge, United Kingdom).

Techniques: Expressing, In Vivo, Isolation, Staining, Injection, Phospho-proteomics, Western Blot, Concentration Assay, Comparison, Control

Journal: Frontiers in Pharmacology

Article Title: Ivabradine Ameliorates Cardiac Diastolic Dysfunction in Diabetic Mice Independent of Heart Rate Reduction

doi: 10.3389/fphar.2021.696635

Figure Lengend Snippet: Ivabradine, but not zatebradine, reduces JNK/p38 MAPK activation and the high glucose (HG)-induced increase in fibrogenic protein expression in vitro (A) NRCFs were pretreated with or without ivabradine at the indicated concentrations for 30 min, and then exposed to HG for 48 h. JNK and p38 MAPK phosphorylation and total JNK and p38 MAPK expression were determined by western blotting (B) NRCFs were treated as described in (A) and the protein expression of PCNA, α-SMA, TIMP2, collagen I, collagen III, and MMP2 was determined by western blotting (C and D) NRCFs were pretreated with or without zatebradine at the indicated concentrations for 30 min, and then exposed to HG for 48 h. The proteins described in (A) and (B) were quantified by western blotting (E and F) NRCFs were treated as described in (A) and (C) and the cell proliferation rate was determined using an MTT assay. Data are presented as mean ± SEM ( n = 3). p -values were calculated using one-way ANOVA with Tukey multiple comparison test. * p < 0.05, compared to the normal group; # p < 0.05, compared to the HG group.

Article Snippet: Antibodies against collagen I and collagen III were obtained from the Biorbyt Corporation (Orwell Furlong, Cambridge, United Kingdom).

Techniques: Activation Assay, Expressing, In Vitro, Phospho-proteomics, Western Blot, MTT Assay, Comparison

Journal: Frontiers in Pharmacology

Article Title: Ivabradine Ameliorates Cardiac Diastolic Dysfunction in Diabetic Mice Independent of Heart Rate Reduction

doi: 10.3389/fphar.2021.696635

Figure Lengend Snippet: Ivabradine, but not zatebradine, ameliorates the diabetes-induced increases in fibrogenic protein expression and fibrosis (A) Collagen I, collagen III, TIMP2, and MMP2 protein expression was determined by western blotting in left ventricular fibroblasts isolated from mice administered ivabradine at the indicated doses (B) Left ventricular fibroblasts were isolated from mice administered zatebradine at the indicated doses and the expression of the proteins listed in (A) was measured by western blotting (C and D) Mice were treated as described in (A) and (B) and the degree of cardiac fibrosis was determined by Massonʼs trichrome staining and immunohistochemistry. The columns show the differences in collagen accumulation (original magnification ×400). Scale bars, 50 μm. Data in (A–B) ( n = 3) and (C–D) ( n = 6) are presented as mean ± SEM. p -values were calculated using one-way ANOVA with Tukey multiple comparison test. * p < 0.05, compared to the Control group; # p < 0.05, compared to the db/db mouse group.

Article Snippet: Antibodies against collagen I and collagen III were obtained from the Biorbyt Corporation (Orwell Furlong, Cambridge, United Kingdom).

Techniques: Expressing, Western Blot, Isolation, Staining, Immunohistochemistry, Comparison, Control

Journal: Acta pharmacologica Sinica

Article Title: Synthesis of the novel PARP-1 inhibitor AG-690/11026014 and its protective effects on angiotensin II-induced mouse cardiac remodeling.

doi: 10.1038/aps.2016.159

Figure Lengend Snippet: Figure 2. 6014 attenuated the enlargement of the heart size, cardiomyocytes size and prevented the accumulation of collagen. (A) Representative diagram of mice hearts under different treatment. (B) Representative images diagram of H&E staining of cardiomyocytes. (C) Representative images of Masson’s staining of total collagen in tissue sections of heart. (D) Statistical results of HW/BW. (E) Statistical results of LVW/BW. (F) Statistical areas of total collagen production versus areas of whole tissue. Data were expressed as mean±SEM from three independent experiments. **P<0.01 vs Sham. #P<0.05, ##P<0.01 vs Ang II.

Article Snippet: Anti-PARP-1, anticollagen I and anti-collagen III antibodies were provided by Proteintech Group (Chicago, IL, USA).

Techniques: Staining

Journal: Acta pharmacologica Sinica

Article Title: Synthesis of the novel PARP-1 inhibitor AG-690/11026014 and its protective effects on angiotensin II-induced mouse cardiac remodeling.

doi: 10.1038/aps.2016.159

Figure Lengend Snippet: Figure 3. Effects of 6014 on the expression of hypertrophy marker proteins and fibrosis marker proteins. Protein expression of ANF (A), BNP (B), β-MHC (C), FN (D), collagen I (E) and collagen III (F) were measured by Western blot. Data were expressed as mean±SEM from duplicates in three independent experiments. **P<0.01 vs Sham. #P<0.05, ##P<0.01 vs Ang II.

Article Snippet: Anti-PARP-1, anticollagen I and anti-collagen III antibodies were provided by Proteintech Group (Chicago, IL, USA).

Techniques: Expressing, Marker, Western Blot

Journal: Acta pharmacologica Sinica

Article Title: Synthesis of the novel PARP-1 inhibitor AG-690/11026014 and its protective effects on angiotensin II-induced mouse cardiac remodeling.

doi: 10.1038/aps.2016.159

Figure Lengend Snippet: Figure 4. Effects of 6014 on the expression and enzymatic activity of PARP-1. The expression of PARP-1 in nucleus (A) and whole cell (B) as well as PARylated proteins in whole cell (C). Data were expressed as mean±SEM from duplicates in three independent experiments. *P<0.05, **P<0.01 vs Sham. ##P<0.01 vs Ang II.

Article Snippet: Anti-PARP-1, anticollagen I and anti-collagen III antibodies were provided by Proteintech Group (Chicago, IL, USA).

Techniques: Expressing, Activity Assay

Journal: Acta pharmacologica Sinica

Article Title: Synthesis of the novel PARP-1 inhibitor AG-690/11026014 and its protective effects on angiotensin II-induced mouse cardiac remodeling.

doi: 10.1038/aps.2016.159

Figure Lengend Snippet: Figure 6. Effects of 6014 on the regulation and activity of SIRT-1. NAD+ content analysis (A) and SIRT-1 activity (B) were analyzed. Protein expression of SIRT-1 were measured by Western blot (C). The interaction between SIRT-1 and PARP-1 (D) and the PARylation of SIRT-1 (E) were measured by co-IP assay. Data were expressed as mean±SEM from three independent experiments. *P<0.05, **P<0.01 vs Sham. #P<0.05, ##P<0.01 vs Ang II.

Article Snippet: Anti-PARP-1, anticollagen I and anti-collagen III antibodies were provided by Proteintech Group (Chicago, IL, USA).

Techniques: Activity Assay, Expressing, Western Blot, Co-Immunoprecipitation Assay

Journal: Current Protocols

Article Title: Cryopreservation of Human Adult Ventricular Tissue for the Preparation of Viable Myocardial Slices

doi: 10.1002/cpz1.70068

Figure Lengend Snippet: Primary Antibodies

Article Snippet: Type III Collagen , 1:2000 , Southern Biotech , 1330‐01.

Techniques:

Journal: Current Protocols

Article Title: Cryopreservation of Human Adult Ventricular Tissue for the Preparation of Viable Myocardial Slices

doi: 10.1002/cpz1.70068

Figure Lengend Snippet: Extracellular matrix. Representative images of immunohistochemical analysis of fresh and cryopreserved cardiac tissue. Type I Collagen (Collagen‐I) is shown in orange (A) and Type III Collagen (Collagen‐III) is shown in yellow (B) . Counterstaining of nuclei was performed with DAPI (blue).

Article Snippet: Type III Collagen , 1:2000 , Southern Biotech , 1330‐01.

Techniques: Immunohistochemical staining

Journal:

Article Title: A mutant, noninhibitory plasminogen activator inhibitor type 1 decreases matrix accumulation in experimental glomerulonephritis

doi: 10.1172/JCI200318038

Figure Lengend Snippet: Immunofluorescent staining score for ECM proteins at d6. Glomerular staining for FN-EDA+ (a), laminin (b), type I collagen (c), and type III collagen (d) were lower in the PAI-1R–treated, nephritic group. *P < 0.001 compared with normal control. #P < 0.01 compared with disease control.

Article Snippet: Monoclonal mouse anti-cellular fibronectin extra domain positive (FN-EDA+) (Harlan Sera-Lab Ltd., Loughborough, United Kingdom), rabbit anti–mouse laminin (ICN Immunobiologicals, Aurora, Ohio, USA), goat anti–human type I collagen, and goat anti–human type III collagen (Southern Biotechnology Associates, Birmingham, Alabama, USA) were used as the primary Ab’s for detection of matrix components in glomeruli at d6.

Techniques: Staining