Journal: Life Science Alliance

Article Title: Local tissue mechanics control cardiac pacemaker cell embryonic patterning

doi: 10.26508/lsa.202201799

Figure Lengend Snippet: (A) Col3A1 staining at the SAN/atria junction in representative HH18, HH30, and HH35 hearts. (B) Distribution of Col3A1, TnC, and WFA in the H35 SAN and atria. (C) Diagram of in vivo transfection strategy for mosaic calcium imaging in the intact SAN. Integrating DNA plasmids were microinjected into the pericardial space adjacent to the forming SAN at HH18 and SAN tissue was isolated for live imaging at HH30. (D) Live imaging of a memRFP/GCaMP6f-positive cell within the SAN. Data presented in were taken from cells imaged in 25 individual hearts.

Article Snippet: The following reagents were used for immunohistochemistry: MF20 (14650380; Invitrogen) (1:500); FLRT3 (PA563240; Invitrogen) (1:250); hyaluronic acid binding protein, bovine nasal cartilage, biotinylated (38591150UG; MilliporeSigma) (1:100); Wisteria floribunda lectin biotinylated (B13552; Vector Laboratories) (1:100); Col3A1 (3B2-s; DSHB) (1:250), TNC (LS-C39574-50; LSBio) (1:500); Nav1.5 (LS-C30483; LSBio) (1:500), phalloidin-647 (A22287; Invitrogen) (1:40); NCX1 (LS-C73201-100; LSBio) (1:500); streptavidin-568 (S11226; Invitrogen) (1:500); DAPI (62248; Thermo Fisher Scientific) (1:1,000); IgM (heavy chain) cross-adsorbed goat anti-mouse, Alexa Fluor 555 (A21426; Invitrogen) (1:500); IgG1 cross-adsorbed goat anti-mouse, Alexa Fluor 647 (A21240; Invitrogen) (1:500); IgG2b cross-adsorbed goat anti-mouse, Alexa Fluor 488 (A21141; Invitrogen) (1:500); IgG (H+L) cross-adsorbed goat anti-rabbit, Alexa Fluor 568 (A11011; Invitrogen) (1:500); goat anti-mouse IgG2b cross-adsorbed secondary antibody, Alexa Fluor 647 (A21242; Invitrogen) (1:500); goat anti-rabbit IgG (H+L) cross-adsorbed secondary antibody, Alexa Fluor 488 (A11008; Invitrogen) (1:500); goat anti-mouse IgM mu chain Alexa Fluor 488 (ab150121; Abcam) (1:500).

Techniques: Staining, In Vivo, Transfection, Imaging, Isolation

Journal: Acta biochimica et biophysica Sinica

Article Title: TXNIP aggravates cardiac fibrosis and dysfunction after myocardial infarction in mice by enhancing the TGFB1/Smad3 pathway and promoting NLRP3 inflammasome activation.

doi: 10.3724/abbs.2023150

Figure Lengend Snippet: Figure 3. TXNIP promoted post-MI collagen deposition (A,B) Representative picrosirius red staining images of heart sections from each group at 28 days after treatment (A) under a light microscope and (B) under a polarized light microscope. Collagen I showed red/orange birefringence, while Collagen III showed green/whitish birefringence. (C,E,G) Representative immunohistochemical staining images of heart sections from each group at 28 days after treatment. (C) Collagen I deposition. (E) Collagen III deposition, and (G) ACTA2 deposition. (D) Statistical analysis of Collagen I deposition. Data are shown as the mean±SEM, n=3. **P<0.01. (F) Statistical analysis of Collagen III deposition. Data are shown as the mean± SEM, n=3. **P<0.01. (H) Statistical analysis of ACTA2 deposition. Data are shown as the mean±SEM, n=3. *P<0.05, **P<0.01.

Article Snippet: Then, the membrane was incubated with one of the following antibodies at 4°C overnight: rabbit antiCollagen I, ACTA2, CTGF, TXNIP, Smad3, p-Smad3 (Abcam), antiSmad7, NLRP3, ASC, Caspase-1/Cleaved Caspase-1, Mature IL1B, GAPDH (Wanleibio, Shenyang, China), mouse anti-TGFB1 antibody (Santa Cruz Biotechnology, Dallas, USA), and mouse anti-Collagen III antibody (Boster).

Techniques: Staining, Light Microscopy, Immunohistochemical staining

Journal: Acta biochimica et biophysica Sinica

Article Title: TXNIP aggravates cardiac fibrosis and dysfunction after myocardial infarction in mice by enhancing the TGFB1/Smad3 pathway and promoting NLRP3 inflammasome activation.

doi: 10.3724/abbs.2023150

Figure Lengend Snippet: Figure 4. TXNIP increased post-MI collagen deposition (A) Representative images of the protein levels of Collagen III, Collagen I, ACTA2, and CTGF. (B‒E) Statistical analyses of the protein levels of Collagen I, Collagen III, ACTA2, and CTGF. Data are shown as the mean±SEM, n=3‒5. *P<0.05, **P<0.01; ns, not significant.

Article Snippet: Then, the membrane was incubated with one of the following antibodies at 4°C overnight: rabbit antiCollagen I, ACTA2, CTGF, TXNIP, Smad3, p-Smad3 (Abcam), antiSmad7, NLRP3, ASC, Caspase-1/Cleaved Caspase-1, Mature IL1B, GAPDH (Wanleibio, Shenyang, China), mouse anti-TGFB1 antibody (Santa Cruz Biotechnology, Dallas, USA), and mouse anti-Collagen III antibody (Boster).

Techniques:

Journal: RSC Advances

Article Title: Thymoquinone protects against cardiac damage from doxorubicin-induced heart failure in Sprague-Dawley rats

doi: 10.1039/c8ra00975a

Figure Lengend Snippet: Fig. 4 Protein levels of TGF-b, Smad3, collagen I, collagen III, and a-SMA in cardiac tissue were determined by western blotting. Immunoblotting for TGF-b, Smad3, collagen I, collagen III, and a-SMA in cardiac tissues. Bar graph showing quantification of TGF-b, Smad3, collagen I, collagen III, and a-SMA protein expression. Data are given as the mean SEM; n ¼ 3 in each group. *p < 0.05 vs. DOX group.

Article Snippet: Primary antibodies against TGF-b (rabbit anti-TGF-b antibody, 1 : 1000; Proteintech, Wuhan, China), Smad3 (rabbit anti-Smad antibody, 1 : 1000; Proteintech), collagen I (rabbit anti-collagen I antibody, 1 : 1000; Proteintech), collagen III (rabbit anticollagen III antibody, 1 : 1000; Proteintech), a-SMA (rabbit anti-a-SMA antibody, 1 : 1000; Proteintech), P53 (rabbit antiP53 antibody, 1 : 1000; Proteintech), bcl-2(rabbit anti-bcl-2 antibody, 1 : 1000; Proteintech), anti-b-actin (1 : 1000; Cell Signaling Technology).

Techniques: Western Blot, Expressing

Journal: Frontiers in Oncology

Article Title: Modeling extracellular matrix through histo-molecular gradient in NSCLC for clinical decisions

doi: 10.3389/fonc.2022.1042766

Figure Lengend Snippet: Violin plots graphs showing proteins of the epithelial-mesenchymal transition process, matricellular, and the Wnt signaling pathway expression analyzed by QuPath (N=120). The protein expression was demonstrated between three different histologic types in non-small cells lung cancer for (A) E-cadherin, (B) β-catenin, (C) Heparan sulfate, (D) Chondroitin sulfate, (E) Col I, (F) Col III, (G) Col IV, (H) Col V, (I) WNT1, (J) WNT3A, (K) WNT5A, (L) WNT5B, and (M) SPARC. LCC, large cell carcinoma; ADC, lung adenocarcinoma; SqCC, lung squamous cell carcinoma; Col, collagen type.

Article Snippet: The specimens were incubated overnight at 4°C with antibodies against: E-cadherin (1:100; Boster Biological), β-catenin (1:100; Santa Cruz), heparan sulfate (1:500; Santa Cruz), chondroitin sulfate (1:100, Santa Cruz), anti-human collagen type I (1:700; Rockland Inc.), anti-human collagen type III (1:200; Rockland Inc.), anti-human collagen type IV (1:100; Dako), and anti-human collagen type V (1:1000; Rockland Inc.).

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: Modeling extracellular matrix through histo-molecular gradient in NSCLC for clinical decisions

doi: 10.3389/fonc.2022.1042766

Figure Lengend Snippet: Co-analysis of immunofluorescence of chondroitin sulfate (green; C , G , K ) and collagen type I (red; B , F , J ) in three different histological subtypes of non-small cell lung carcinoma (N=120). The stained nuclei are represented in blue (DAPI; A , E , I ). Images D , H , L represent the merge of the same field of these three stains. White arrows indicate positive expression of the markers. Original magnification: 40x. LCC, large cell carcinoma; ADC, lung adenocarcinoma; SqCC: lung squamous cell carcinoma.

Article Snippet: The specimens were incubated overnight at 4°C with antibodies against: E-cadherin (1:100; Boster Biological), β-catenin (1:100; Santa Cruz), heparan sulfate (1:500; Santa Cruz), chondroitin sulfate (1:100, Santa Cruz), anti-human collagen type I (1:700; Rockland Inc.), anti-human collagen type III (1:200; Rockland Inc.), anti-human collagen type IV (1:100; Dako), and anti-human collagen type V (1:1000; Rockland Inc.).

Techniques: Immunofluorescence, Staining, Expressing

Journal: Frontiers in Oncology

Article Title: Modeling extracellular matrix through histo-molecular gradient in NSCLC for clinical decisions

doi: 10.3389/fonc.2022.1042766

Figure Lengend Snippet: Co-analysis of immunofluorescence of chondroitin sulfate (green; C , G , K ) and collagen type V (red; B , F , J ) in three different histological subtypes of non-small cell lung carcinoma (N=120). The stained nuclei are represented in blue (DAPI; A , E , I ). Images D , H , L represent the merge of the same field of these three stains. White arrows indicate positive expression of the markers. Original magnification: 40x. LCC, large cell carcinoma; ADC, lung adenocarcinoma; SqCC: lung squamous cell carcinoma.

Article Snippet: The specimens were incubated overnight at 4°C with antibodies against: E-cadherin (1:100; Boster Biological), β-catenin (1:100; Santa Cruz), heparan sulfate (1:500; Santa Cruz), chondroitin sulfate (1:100, Santa Cruz), anti-human collagen type I (1:700; Rockland Inc.), anti-human collagen type III (1:200; Rockland Inc.), anti-human collagen type IV (1:100; Dako), and anti-human collagen type V (1:1000; Rockland Inc.).

Techniques: Immunofluorescence, Staining, Expressing

Journal: Frontiers in Oncology

Article Title: Modeling extracellular matrix through histo-molecular gradient in NSCLC for clinical decisions

doi: 10.3389/fonc.2022.1042766

Figure Lengend Snippet: Co-analysis of immunofluorescence of chondroitin sulfate (green; C , G , K ) and collagen type III (red; B , F , J ) in three different histological subtypes of non-small cell lung carcinoma (N=120). The stained nuclei are represented in blue (DAPI; A , E , I ). Images D , H , L represent the merge of the same field of these three stains. White arrows indicate positive expression of the markers. Original magnification: 40x. LCC, large cell carcinoma; ADC, lung adenocarcinoma; SqCC: lung squamous cell carcinoma.

Article Snippet: The specimens were incubated overnight at 4°C with antibodies against: E-cadherin (1:100; Boster Biological), β-catenin (1:100; Santa Cruz), heparan sulfate (1:500; Santa Cruz), chondroitin sulfate (1:100, Santa Cruz), anti-human collagen type I (1:700; Rockland Inc.), anti-human collagen type III (1:200; Rockland Inc.), anti-human collagen type IV (1:100; Dako), and anti-human collagen type V (1:1000; Rockland Inc.).

Techniques: Immunofluorescence, Staining, Expressing

Journal: Frontiers in Oncology

Article Title: Modeling extracellular matrix through histo-molecular gradient in NSCLC for clinical decisions

doi: 10.3389/fonc.2022.1042766

Figure Lengend Snippet: Co-analysis of immunofluorescence of heparan sulfate (green; C , G , K ) and collagen type IV (red; B , F , J ) in three different histological subtypes of non-small cell lung carcinoma (N=120). The stained nuclei are represented in blue (DAPI; A , E , I ). Images D , H , L represent the merge of the same field of these three stains. White arrows indicate positive expression of the markers. Original magnification: 40x. LCC, large cell carcinoma; ADC, lung adenocarcinoma; SqCC: lung squamous cell carcinoma.

Article Snippet: The specimens were incubated overnight at 4°C with antibodies against: E-cadherin (1:100; Boster Biological), β-catenin (1:100; Santa Cruz), heparan sulfate (1:500; Santa Cruz), chondroitin sulfate (1:100, Santa Cruz), anti-human collagen type I (1:700; Rockland Inc.), anti-human collagen type III (1:200; Rockland Inc.), anti-human collagen type IV (1:100; Dako), and anti-human collagen type V (1:1000; Rockland Inc.).

Techniques: Immunofluorescence, Staining, Expressing

Journal: Frontiers in Oncology

Article Title: Modeling extracellular matrix through histo-molecular gradient in NSCLC for clinical decisions

doi: 10.3389/fonc.2022.1042766

Figure Lengend Snippet: Association between clinicopathologic characteristics and mean expression (% positive expression) of GAGs and collagen types ( t test and ANOVA, P <0.05).

Article Snippet: The specimens were incubated overnight at 4°C with antibodies against: E-cadherin (1:100; Boster Biological), β-catenin (1:100; Santa Cruz), heparan sulfate (1:500; Santa Cruz), chondroitin sulfate (1:100, Santa Cruz), anti-human collagen type I (1:700; Rockland Inc.), anti-human collagen type III (1:200; Rockland Inc.), anti-human collagen type IV (1:100; Dako), and anti-human collagen type V (1:1000; Rockland Inc.).

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: Modeling extracellular matrix through histo-molecular gradient in NSCLC for clinical decisions

doi: 10.3389/fonc.2022.1042766

Figure Lengend Snippet: Variables associated with overall survival in 120 non-small cell lung cancer patients.

Article Snippet: The specimens were incubated overnight at 4°C with antibodies against: E-cadherin (1:100; Boster Biological), β-catenin (1:100; Santa Cruz), heparan sulfate (1:500; Santa Cruz), chondroitin sulfate (1:100, Santa Cruz), anti-human collagen type I (1:700; Rockland Inc.), anti-human collagen type III (1:200; Rockland Inc.), anti-human collagen type IV (1:100; Dako), and anti-human collagen type V (1:1000; Rockland Inc.).

Techniques: Expressing

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: Journal of Diabetes Research

Article Title: Relationship between Urinary Liver-Type Fatty Acid-Binding Protein (L-FABP) and Sarcopenia in Spontaneously Diabetic Torii Fatty Rats

doi: 10.1155/2020/7614035

Figure Lengend Snippet: Histological evaluation of the kidney in SD and SDT fatty rats. Immunohistological staining of kidney tissues using antibodies against CD68 (a, b), α -SMA (c, d), type I collagen (e, f), and type III collagen (g, h). Histological PAS staining showing focal glomerular sclerosis (i) and semiquantitative assessment of focal glomerular sclerosis (j). Original magnification: ×100. Values are presented as the mean ± SEM. ∗ p < 0.05 vs. SD rats.

Article Snippet: The tissue specimens fixed in methyl Carnoy's solution were assessed immunohistochemically for macrophages using a mouse monoclonal antibody (ED-1) specific for CD68 (1 : 100; Abcam, Tokyo, Japan) and goat polyclonal antibodies specific for type I and III collagen (1 : 200; Southern Biotech, Birmingham, AL, USA).

Techniques: Staining

Journal: Journal of Diabetes Research

Article Title: Relationship between Urinary Liver-Type Fatty Acid-Binding Protein (L-FABP) and Sarcopenia in Spontaneously Diabetic Torii Fatty Rats

doi: 10.1155/2020/7614035

Figure Lengend Snippet: Relationships between renal morphological change and muscle strength, and weights of the soleus and extensor digitorum longus (EDL) muscles in 24-week-old SD and SDT fatty rats.

Article Snippet: The tissue specimens fixed in methyl Carnoy's solution were assessed immunohistochemically for macrophages using a mouse monoclonal antibody (ED-1) specific for CD68 (1 : 100; Abcam, Tokyo, Japan) and goat polyclonal antibodies specific for type I and III collagen (1 : 200; Southern Biotech, Birmingham, AL, USA).

Techniques: Muscles, Expressing

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: Experimental & molecular medicine

Article Title: Loss of KDM5B ameliorates pathological cardiac fibrosis and dysfunction by epigenetically enhancing ATF3 expression.

doi: 10.1038/s12276-022-00904-y

Figure Lengend Snippet: Fig. 2 KDM5B deficiency protects the heart from dysfunction and adverse cardiac remodeling after MI. a Echocardiographic measurement of the LVEF, LVFS, left ventricular end-diastolic internal dimension (LVIDd), and left ventricular end-systolic internal dimension (LVIDs) of KDM5B-KO or WT mice at baseline (Day 0) and on the indicated day after MI or sham operation (n = 6 mice per group). b–e Representative Masson’s trichrome staining images and quantitation of the scar size (b, c) or Sirius red staining images and quantitation of the fibrosis area (d, e) in myocardial tissues from KDM5B-KO or WT mice on Day 28 after MI. n = 6 mice per group. Scale bar, 1.6 mm (upper), 200 μm (bottom). f Q-PCR analysis of Col1a1 and Col3a1 mRNA levels in myocardial tissues from KDM5B-KO or WT mice on Day 14 after MI or sham operation (n = 6 mice per group). g Representative immunofluorescence staining of α-SMA (red) and Col III (green) in myocardial tissues from KDM5B-KO or WT mice on Day 14 after MI (n = 6 mice per group). Scale bar, 50 μm. *p < 0.05, **p < 0.01, ***p < 0.001. Unpaired Student’s t test (c, e) or ANOVA (a, f) was performed.

Article Snippet: Antibodies against collagen type III (NB600-594) were obtained from Novus Biologicals (Littleton, CO).

Techniques: Staining, Quantitation Assay

Journal: Experimental & molecular medicine

Article Title: Loss of KDM5B ameliorates pathological cardiac fibrosis and dysfunction by epigenetically enhancing ATF3 expression.

doi: 10.1038/s12276-022-00904-y

Figure Lengend Snippet: Fig. 3 KDM5B deficiency prevents pressure overload-induced cardiac dysfunction and cardiac fibrosis. a Representative echocardio- graphic M-mode images of left ventricles from KDM5B-KO or littermate control WT mice on Day 28 after AngII or normal saline (NS) infusion. b, c Echocardiographic measurement of the LVEF (b) and LVFS (c) of KDM5B-KO or WT mice on Day 28 after AngII or NS infusion (n = 6 mice per group). d, e The ratio of heart weight to body weight (HW/BW) (d) and the ratio of heart weight to tibia length (HW/TL) (e) of KDM5B-KO or WT mice on Day 28 after AngII or NS infusion (n = 6 mice per group). f–i Representative Masson’s trichrome images and quantitation of perivascular (f, g) or interstitial (h, i) fibrosis in myocardial tissues from KDM5B-KO or WT mice on Day 28 after AngII or NS infusion (n = 6 mice per group). Scale bar, 200 μm (left), 100 μm (right). j Q-PCR analysis of Acta2, Col1a1, Col3a1 and Ctgf mRNA expression levels in myocardial tissues from KDM5B-KO or WT mice on Day 28 after AngII or NS infusion (n = 6 mice per group). k Immunoblot analysis of Col I and Col III protein expression in myocardial tissues from KDM5B-KO or WT mice on Day 28 after AngII or NS infusion. l Representative immunofluorescence staining of α-SMA (red) and Col III (green) in myocardial tissues from KDM5B-KO or WT mice on Day 28 after AngII infusion. Scale bar, 50 μm. *p < 0.05, **p < 0.01. Unpaired Student’s t test (g, i) or one-way ANOVA (b–e, j) was performed.

Article Snippet: Antibodies against collagen type III (NB600-594) were obtained from Novus Biologicals (Littleton, CO).

Techniques: Control, Saline, Quantitation Assay, Expressing, Western Blot, Staining

Journal: Experimental & molecular medicine

Article Title: Loss of KDM5B ameliorates pathological cardiac fibrosis and dysfunction by epigenetically enhancing ATF3 expression.

doi: 10.1038/s12276-022-00904-y

Figure Lengend Snippet: Fig. 4 KDM5B promotes fibrotic responses and the transition of cardiac fibroblasts. a, b Q-PCR analysis of Col1a1, Col3a1, Fn1 and Ccn2 mRNA expression (a) (n = 6 per group) or immunoblot analysis of Col I and Col III protein expression (b) in KDM5B-deficient (KO) or littermate control WT cardiac fibroblasts stimulated with TGF-β (10 ng/ml) for 24 h. c, d Q-PCR analysis of Col1a1, Col3a1, Fn1 and Ccn2 mRNA expression (c) (n = 6 per group) or immunoblot analysis of Col I and Col III protein expression (d) in Kdm5b-silenced or control siRNA-transfected cardiac fibroblasts stimulated with TGF-β (10 ng/ml) for 24 h. e, f Q-PCR analysis of Col1a1, Col3a1, Fn1 and Ccn2 mRNA expression (e) (n = 6 per group) or immunoblot analysis of Col I and Col III protein expression (f) in cardiac fibroblasts treated with the KDM5B inhibitor GSK467 or DMSO followed by stimulation with TGF-β (10 ng/ml) for 24 h. g–i Representative immunofluorescence staining of α-SMA (red) in cardiac fibroblasts with KDM5B-deficiency (g), KDM5B knockdown (h) or GSK467 treatment (i) and the corresponding control cardiac fibroblasts (g–i) stimulated with TGF-β (10 ng/ml) for 24 h. Similar results were obtained from three independent experiments (g–i). Scale bar, 50 μm. *p < 0.05, **p < 0.01, ***p < 0.001. Unpaired Student’s t test (a, c, e) was performed.

Article Snippet: Antibodies against collagen type III (NB600-594) were obtained from Novus Biologicals (Littleton, CO).

Techniques: Expressing, Western Blot, Control, Transfection, Staining, Knockdown