Journal: Journal of Sport and Health Science

Article Title: Influence of diet-induced obesity and voluntary exercise training on cardiac lipids and mitochondrial function in mice

doi: 10.1016/j.jshs.2025.101095

Figure Lengend Snippet: Impact of high fat feeding and voluntary exercise on left ventricle morphology in male mice. (A) Representative images of Picrosirius red staining for measurement of interstitial and perivascular fibrosis with 20× magnification. (B) Representative images for Hematoxylin and Eosin for measurement of cardiomyocyte width and area with 40× magnification. (C) Representative images of Oil red O staining for measurement of interstitial lipid deposition with 40× magnification. Quantitative analysis of percent area of (D) interstitial and (E) perivascular fibrosis expressed as fold change from sedentary chow group. Quantitative analysis of (F) cardiomyocyte width and (G) cardiomyocyte area. (H) Quantitative analysis of percent area of lipid deposition expressed as fold change from sedentary chow group left ventricle mRNA expression of fatty acid transporters (I) FABP3, (J) CD36, and hypertrophy markers (K) β-MHC and (L) ANP in male VET or sedentary mice fed an HFD or chow diet. Values were calculated relative to 18S housekeeper. Analysis was performed using two-way analysis of variance with Tukey’s post hoc test for multiple comparisons. Data are expressed as mean ± standard error of the mean. (D–H) n : 8–12 per group. (I–L) n : 8–9 per group. * p < 0.05, **** p < 0.0001. ANP = atrial natriuretic peptide; β-MHC = beta myosin heavy chain; CD36 = platelet glycoprotein 4; FABP3 = fatty acid-binding protein 3; HFD = high fat diet; Myh7 = moysin heavy chain 7; VET = voluntary exercise training.

Article Snippet: The following TaqMan assay gene transcripts were used: fatty acid-binding protein 3 (FABP3, Mm02342495_m1), platelet glycoprotein 4 (CD36, Mm00432403_m1), beta myosin heavy chain (β-MHC, Mm00600555_m1), atrial natriuretic peptide (ANP, Mm01255747_g1), OPA1 (Mm01349707_g1), DRP1 (Mm01342903_m1), robosomal18S (18S, Mm03928990_g1).

Techniques: Staining, Expressing, Binding Assay

Journal: Journal of Sport and Health Science

Article Title: Influence of diet-induced obesity and voluntary exercise training on cardiac lipids and mitochondrial function in mice

doi: 10.1016/j.jshs.2025.101095

Figure Lengend Snippet: Impact of high fat feeding and voluntary exercise on left ventricle morphology in male mice. (A) Representative images of Picrosirius red staining for measurement of interstitial and perivascular fibrosis with 20× magnification. (B) Representative images for Hematoxylin and Eosin for measurement of cardiomyocyte width and area with 40× magnification. (C) Representative images of Oil red O staining for measurement of interstitial lipid deposition with 40× magnification. Quantitative analysis of percent area of (D) interstitial and (E) perivascular fibrosis expressed as fold change from sedentary chow group. Quantitative analysis of (F) cardiomyocyte width and (G) cardiomyocyte area. (H) Quantitative analysis of percent area of lipid deposition expressed as fold change from sedentary chow group left ventricle mRNA expression of fatty acid transporters (I) FABP3, (J) CD36, and hypertrophy markers (K) β-MHC and (L) ANP in male VET or sedentary mice fed an HFD or chow diet. Values were calculated relative to 18S housekeeper. Analysis was performed using two-way analysis of variance with Tukey’s post hoc test for multiple comparisons. Data are expressed as mean ± standard error of the mean. (D–H) n : 8–12 per group. (I–L) n : 8–9 per group. * p < 0.05, **** p < 0.0001. ANP = atrial natriuretic peptide; β-MHC = beta myosin heavy chain; CD36 = platelet glycoprotein 4; FABP3 = fatty acid-binding protein 3; HFD = high fat diet; Myh7 = moysin heavy chain 7; VET = voluntary exercise training.

Article Snippet: The following TaqMan assay gene transcripts were used: fatty acid-binding protein 3 (FABP3, Mm02342495_m1), platelet glycoprotein 4 (CD36, Mm00432403_m1), beta myosin heavy chain (β-MHC, Mm00600555_m1), atrial natriuretic peptide (ANP, Mm01255747_g1), OPA1 (Mm01349707_g1), DRP1 (Mm01342903_m1), robosomal18S (18S, Mm03928990_g1).

Techniques: Staining, Expressing, Binding Assay

Journal: Journal of Sport and Health Science

Article Title: Influence of diet-induced obesity and voluntary exercise training on cardiac lipids and mitochondrial function in mice

doi: 10.1016/j.jshs.2025.101095

Figure Lengend Snippet: Impact of high fat feeding and voluntary exercise on left ventricle morphology in male mice. (A) Representative images of Picrosirius red staining for measurement of interstitial and perivascular fibrosis with 20× magnification. (B) Representative images for Hematoxylin and Eosin for measurement of cardiomyocyte width and area with 40× magnification. (C) Representative images of Oil red O staining for measurement of interstitial lipid deposition with 40× magnification. Quantitative analysis of percent area of (D) interstitial and (E) perivascular fibrosis expressed as fold change from sedentary chow group. Quantitative analysis of (F) cardiomyocyte width and (G) cardiomyocyte area. (H) Quantitative analysis of percent area of lipid deposition expressed as fold change from sedentary chow group left ventricle mRNA expression of fatty acid transporters (I) FABP3, (J) CD36, and hypertrophy markers (K) β-MHC and (L) ANP in male VET or sedentary mice fed an HFD or chow diet. Values were calculated relative to 18S housekeeper. Analysis was performed using two-way analysis of variance with Tukey’s post hoc test for multiple comparisons. Data are expressed as mean ± standard error of the mean. (D–H) n : 8–12 per group. (I–L) n : 8–9 per group. * p < 0.05, **** p < 0.0001. ANP = atrial natriuretic peptide; β-MHC = beta myosin heavy chain; CD36 = platelet glycoprotein 4; FABP3 = fatty acid-binding protein 3; HFD = high fat diet; Myh7 = moysin heavy chain 7; VET = voluntary exercise training.

Article Snippet: The following TaqMan assay gene transcripts were used: fatty acid-binding protein 3 (FABP3, Mm02342495_m1), platelet glycoprotein 4 (CD36, Mm00432403_m1), beta myosin heavy chain (β-MHC, Mm00600555_m1), atrial natriuretic peptide (ANP, Mm01255747_g1), OPA1 (Mm01349707_g1), DRP1 (Mm01342903_m1), robosomal18S (18S, Mm03928990_g1).

Techniques: Staining, Expressing, Binding Assay

Journal: Bioactive Materials

Article Title: Esophagus extracellular matrix with microenvironmental complexity for esophageal organoids

doi: 10.1016/j.bioactmat.2025.12.046

Figure Lengend Snippet: The 3D culture of esophageal organoids in EEM hydrogel. a) Brightfield images of esophageal organoids cultured in various concentrations of EEM hydrogels and MAT at day 9. b) Quantification of organoid formation efficiency in EEM hydrogels, relative to MAT ( n = 3, one-way ANOVA with Tukey's multiple comparisons test; ∗∗ p < 0.01; ns, non-significant difference with p > 0.05). c) qRT-PCR analysis to compare gene expression of a basal layer marker ( Krt14 ) and a suprabasal layer marker ( Krt13 ) in esophageal organoids cultured in EEM hydrogels and MAT at day 9 ( n = 4, one-way ANOVA with Tukey's multiple comparisons test; ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus MAT; ns, non-significant difference with p > 0.05). d) Relative gene expression of basal layer markers ( Sox2 and Krt14 ) and suprabasal layer markers ( Krt13 and Krt4 ) in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT at day 9 ( n = 4; unpaired two-tailed t -test; ∗∗∗ p < 0.001 versus MAT). e) Morphology of esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 9 days. f) Immunofluorescent staining for basal layer markers (CK14, SOX2, and p63), suprabasal layer markers (CK13 and CK4), F-actin, a cell-cell adhesion marker (ECAD), and a proliferation marker (Ki67) in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 12 days. g) Quantification of CK14 and CK13 positive area in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 12 days, based on immunofluorescence-stained images ( n = 4; unpaired two-tailed t -test; ∗ p < 0.05 versus MAT; ns, non-significant difference with p > 0.05). h) Live/Dead staining to assess the viability of organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 11 days. i) Representative brightfield images at passage 0, 2, 4, and 6 of esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT. Total culture days are given in parentheses. j) Comparison of gene expression of a basal layer marker ( Krt14 ) and a suprabasal layer marker ( Krt13 ) in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT at passage 0 and 6 ( n = 4; unpaired two-tailed t -test; ∗∗∗ p < 0.001 versus MAT at each passage).

Article Snippet: The relative gene expression was evaluated using the primers for the following genes (all from Thermo Fisher Scientific): sex-determining region Y-box 2 ( Sox2 ; Mm03053810_s1), keratin 14 ( Krt14 ; Mm00516876_m1), keratin 13 ( Krt13 ; Mm00495199_m1), keratin 4 ( Krt4 ; Mm01296260_m1), keratin 18 ( Krt18 ; Mm01601704_g1), keratin 19 ( Krt19 ; Mm00492980_m1), alpha fetoprotein ( Afp ; Mm00431715_m1), keratin 5 ( Krt5 ; Mm01305291_g1), secretoglobin family 1A member 1 ( Scgb1a1 ; Mm00442046_m1), and forkhead box J1 ( Foxj1 ; Mm01267279_m1).

Techniques: Cell Culture, Quantitative RT-PCR, Gene Expression, Marker, Two Tailed Test, Staining, Immunofluorescence, Comparison

Journal: Bioactive Materials

Article Title: Esophagus extracellular matrix with microenvironmental complexity for esophageal organoids

doi: 10.1016/j.bioactmat.2025.12.046

Figure Lengend Snippet: Tissue-specific effects of EEM on esophageal organoid culture. a) Brightfield images of esophageal organoids cultured for 11 days in various ECM hydrogels (5 mg mL −1 ) produced from different decellularized organs. b) qRT-PCR analysis to compare gene expression of a basal layer marker ( Krt14 ) and a suprabasal layer marker ( Krt13 ) in esophageal organoids cultured in various ECM hydrogels for 11 days ( n = 4; one-way ANOVA with Tukey's multiple comparisons test; ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus EEM; ns, non-significant difference with p > 0.05). c) Representative immunofluorescence staining images showing CK14 (a basal layer marker) and CK13 (a suprabasal layer marker) expression in esophageal organoids cultured in various ECM hydrogels for 11 days. d) Quantification of CK14 + and CK13 + areas in esophageal organoids cultured in various ECM hydrogels for 11 days, based on immunofluorescence staining ( n = 4; one-way ANOVA with Tukey's multiple comparisons test; ∗ p < 0.05 and ∗∗ p < 0.01 versus EEM; ns, non-significant difference with p > 0.05).

Article Snippet: The relative gene expression was evaluated using the primers for the following genes (all from Thermo Fisher Scientific): sex-determining region Y-box 2 ( Sox2 ; Mm03053810_s1), keratin 14 ( Krt14 ; Mm00516876_m1), keratin 13 ( Krt13 ; Mm00495199_m1), keratin 4 ( Krt4 ; Mm01296260_m1), keratin 18 ( Krt18 ; Mm01601704_g1), keratin 19 ( Krt19 ; Mm00492980_m1), alpha fetoprotein ( Afp ; Mm00431715_m1), keratin 5 ( Krt5 ; Mm01305291_g1), secretoglobin family 1A member 1 ( Scgb1a1 ; Mm00442046_m1), and forkhead box J1 ( Foxj1 ; Mm01267279_m1).

Techniques: Cell Culture, Produced, Quantitative RT-PCR, Gene Expression, Marker, Immunofluorescence, Staining, Expressing

Journal: Bioactive Materials

Article Title: Esophagus extracellular matrix with microenvironmental complexity for esophageal organoids

doi: 10.1016/j.bioactmat.2025.12.046

Figure Lengend Snippet: Storage of EEM as the pre-gel solution and its application in organoid culture. Graphical representation for storage of EEM pre-gel solution at a) 4 °C or d) −80 °C. Brightfield images of esophageal organoids cultured in EEM hydrogels formed with pre-gel solution stored at b) 4 °C (for 1 week–1 month) or e, g) −80 °C (for 1 month–2 years), compared to freshly prepared EEM hydrogel and MAT. Relative gene expression of a basal layer marker ( Krt14 ) and suprabasal layer markers ( Krt13 and Krt4 ) in esophageal organoids cultured in EEM hydrogels formed with pre-gel solution stored at c) 4 °C (for 1 week–1 month) or f, h) −80 °C (for 1 month–2 years), compared to freshly prepared EEM hydrogel and MAT ( n = 3–4; one-way ANOVA with Tukey's multiple comparisons test; ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus MAT; ns, non-significant difference with p > 0.05). i) Representative images from immunofluorescence staining of CK14 (a basal layer marker) and CK13 (a suprabasal layer marker) in esophageal organoids cultured in EEM hydrogels formed with pre-gel solution stored at 4 °C or −80 °C for 2 months, compared to freshly prepared EEM hydrogel and MAT. j) Percentages of CK14 + and CK13 + areas relative to the total area in esophageal organoids based on immunofluorescence results shown in (i) ( n = 4; one-way ANOVA with Tukey's multiple comparisons test; ∗∗∗ p < 0.001 versus MAT; ns, non-significant difference with p > 0.05).

Article Snippet: The relative gene expression was evaluated using the primers for the following genes (all from Thermo Fisher Scientific): sex-determining region Y-box 2 ( Sox2 ; Mm03053810_s1), keratin 14 ( Krt14 ; Mm00516876_m1), keratin 13 ( Krt13 ; Mm00495199_m1), keratin 4 ( Krt4 ; Mm01296260_m1), keratin 18 ( Krt18 ; Mm01601704_g1), keratin 19 ( Krt19 ; Mm00492980_m1), alpha fetoprotein ( Afp ; Mm00431715_m1), keratin 5 ( Krt5 ; Mm01305291_g1), secretoglobin family 1A member 1 ( Scgb1a1 ; Mm00442046_m1), and forkhead box J1 ( Foxj1 ; Mm01267279_m1).

Techniques: Cell Culture, Gene Expression, Marker, Immunofluorescence, Staining

Journal: Bioactive Materials

Article Title: Esophagus extracellular matrix with microenvironmental complexity for esophageal organoids

doi: 10.1016/j.bioactmat.2025.12.046

Figure Lengend Snippet: The 3D culture of esophageal organoids in EEM hydrogel. a) Brightfield images of esophageal organoids cultured in various concentrations of EEM hydrogels and MAT at day 9. b) Quantification of organoid formation efficiency in EEM hydrogels, relative to MAT ( n = 3, one-way ANOVA with Tukey's multiple comparisons test; ∗∗ p < 0.01; ns, non-significant difference with p > 0.05). c) qRT-PCR analysis to compare gene expression of a basal layer marker ( Krt14 ) and a suprabasal layer marker ( Krt13 ) in esophageal organoids cultured in EEM hydrogels and MAT at day 9 ( n = 4, one-way ANOVA with Tukey's multiple comparisons test; ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus MAT; ns, non-significant difference with p > 0.05). d) Relative gene expression of basal layer markers ( Sox2 and Krt14 ) and suprabasal layer markers ( Krt13 and Krt4 ) in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT at day 9 ( n = 4; unpaired two-tailed t -test; ∗∗∗ p < 0.001 versus MAT). e) Morphology of esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 9 days. f) Immunofluorescent staining for basal layer markers (CK14, SOX2, and p63), suprabasal layer markers (CK13 and CK4), F-actin, a cell-cell adhesion marker (ECAD), and a proliferation marker (Ki67) in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 12 days. g) Quantification of CK14 and CK13 positive area in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 12 days, based on immunofluorescence-stained images ( n = 4; unpaired two-tailed t -test; ∗ p < 0.05 versus MAT; ns, non-significant difference with p > 0.05). h) Live/Dead staining to assess the viability of organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT for 11 days. i) Representative brightfield images at passage 0, 2, 4, and 6 of esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT. Total culture days are given in parentheses. j) Comparison of gene expression of a basal layer marker ( Krt14 ) and a suprabasal layer marker ( Krt13 ) in esophageal organoids cultured in EEM hydrogel (5 mg mL −1 ) and MAT at passage 0 and 6 ( n = 4; unpaired two-tailed t -test; ∗∗∗ p < 0.001 versus MAT at each passage).

Article Snippet: The relative gene expression was evaluated using the primers for the following genes (all from Thermo Fisher Scientific): sex-determining region Y-box 2 ( Sox2 ; Mm03053810_s1), keratin 14 ( Krt14 ; Mm00516876_m1), keratin 13 ( Krt13 ; Mm00495199_m1), keratin 4 ( Krt4 ; Mm01296260_m1), keratin 18 ( Krt18 ; Mm01601704_g1), keratin 19 ( Krt19 ; Mm00492980_m1), alpha fetoprotein ( Afp ; Mm00431715_m1), keratin 5 ( Krt5 ; Mm01305291_g1), secretoglobin family 1A member 1 ( Scgb1a1 ; Mm00442046_m1), and forkhead box J1 ( Foxj1 ; Mm01267279_m1).

Techniques: Cell Culture, Quantitative RT-PCR, Gene Expression, Marker, Two Tailed Test, Staining, Immunofluorescence, Comparison

Journal: Bioactive Materials

Article Title: Esophagus extracellular matrix with microenvironmental complexity for esophageal organoids

doi: 10.1016/j.bioactmat.2025.12.046

Figure Lengend Snippet: Tissue-specific effects of EEM on esophageal organoid culture. a) Brightfield images of esophageal organoids cultured for 11 days in various ECM hydrogels (5 mg mL −1 ) produced from different decellularized organs. b) qRT-PCR analysis to compare gene expression of a basal layer marker ( Krt14 ) and a suprabasal layer marker ( Krt13 ) in esophageal organoids cultured in various ECM hydrogels for 11 days ( n = 4; one-way ANOVA with Tukey's multiple comparisons test; ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus EEM; ns, non-significant difference with p > 0.05). c) Representative immunofluorescence staining images showing CK14 (a basal layer marker) and CK13 (a suprabasal layer marker) expression in esophageal organoids cultured in various ECM hydrogels for 11 days. d) Quantification of CK14 + and CK13 + areas in esophageal organoids cultured in various ECM hydrogels for 11 days, based on immunofluorescence staining ( n = 4; one-way ANOVA with Tukey's multiple comparisons test; ∗ p < 0.05 and ∗∗ p < 0.01 versus EEM; ns, non-significant difference with p > 0.05).

Article Snippet: The relative gene expression was evaluated using the primers for the following genes (all from Thermo Fisher Scientific): sex-determining region Y-box 2 ( Sox2 ; Mm03053810_s1), keratin 14 ( Krt14 ; Mm00516876_m1), keratin 13 ( Krt13 ; Mm00495199_m1), keratin 4 ( Krt4 ; Mm01296260_m1), keratin 18 ( Krt18 ; Mm01601704_g1), keratin 19 ( Krt19 ; Mm00492980_m1), alpha fetoprotein ( Afp ; Mm00431715_m1), keratin 5 ( Krt5 ; Mm01305291_g1), secretoglobin family 1A member 1 ( Scgb1a1 ; Mm00442046_m1), and forkhead box J1 ( Foxj1 ; Mm01267279_m1).

Techniques: Cell Culture, Produced, Quantitative RT-PCR, Gene Expression, Marker, Immunofluorescence, Staining, Expressing

Journal: Bioactive Materials

Article Title: Esophagus extracellular matrix with microenvironmental complexity for esophageal organoids

doi: 10.1016/j.bioactmat.2025.12.046

Figure Lengend Snippet: Storage of EEM as the pre-gel solution and its application in organoid culture. Graphical representation for storage of EEM pre-gel solution at a) 4 °C or d) −80 °C. Brightfield images of esophageal organoids cultured in EEM hydrogels formed with pre-gel solution stored at b) 4 °C (for 1 week–1 month) or e, g) −80 °C (for 1 month–2 years), compared to freshly prepared EEM hydrogel and MAT. Relative gene expression of a basal layer marker ( Krt14 ) and suprabasal layer markers ( Krt13 and Krt4 ) in esophageal organoids cultured in EEM hydrogels formed with pre-gel solution stored at c) 4 °C (for 1 week–1 month) or f, h) −80 °C (for 1 month–2 years), compared to freshly prepared EEM hydrogel and MAT ( n = 3–4; one-way ANOVA with Tukey's multiple comparisons test; ∗ p < 0.05, ∗∗ p < 0.01, and ∗∗∗ p < 0.001 versus MAT; ns, non-significant difference with p > 0.05). i) Representative images from immunofluorescence staining of CK14 (a basal layer marker) and CK13 (a suprabasal layer marker) in esophageal organoids cultured in EEM hydrogels formed with pre-gel solution stored at 4 °C or −80 °C for 2 months, compared to freshly prepared EEM hydrogel and MAT. j) Percentages of CK14 + and CK13 + areas relative to the total area in esophageal organoids based on immunofluorescence results shown in (i) ( n = 4; one-way ANOVA with Tukey's multiple comparisons test; ∗∗∗ p < 0.001 versus MAT; ns, non-significant difference with p > 0.05).

Article Snippet: The relative gene expression was evaluated using the primers for the following genes (all from Thermo Fisher Scientific): sex-determining region Y-box 2 ( Sox2 ; Mm03053810_s1), keratin 14 ( Krt14 ; Mm00516876_m1), keratin 13 ( Krt13 ; Mm00495199_m1), keratin 4 ( Krt4 ; Mm01296260_m1), keratin 18 ( Krt18 ; Mm01601704_g1), keratin 19 ( Krt19 ; Mm00492980_m1), alpha fetoprotein ( Afp ; Mm00431715_m1), keratin 5 ( Krt5 ; Mm01305291_g1), secretoglobin family 1A member 1 ( Scgb1a1 ; Mm00442046_m1), and forkhead box J1 ( Foxj1 ; Mm01267279_m1).

Techniques: Cell Culture, Gene Expression, Marker, Immunofluorescence, Staining