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: Voluntary exercise alters left ventricle gene and protein expression related to mitochondrial biogenesis and dynamics in high fat- and chow-fed mice. (A) Western blot analysis to assess purity of crude mitochondria isolation where left ventricle mitochondrial pellet expresses mitochondrial marker TOM70 and COXIV and supernatant expresses calnexin. Protein expression of (B) LCLAT1 and (C) MFN2 in left ventricle-isolated crude mitochondria and (D) LCLAT1, (E) PGC-1α, and (F) MFN2 in left ventricle tissue from male VET or sedentary mice fed an HFD or chow diet. Left ventricle mRNA expression of (G) OPA1 and (H) DRP1 in male VET or sedentary mice fed an HFD or chow diet. Values were calculated relative to 18S housekeeper. Calnexin and β-actin were used as internal controls of protein loading in left ventricle samples and COXIV as internal control of protein loading in crude mitochondria samples. 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. (B–C) n : 5–6 per group. (D–F) n = 7 per group. (G and H) n = 9 per group. CE = chow exercise; CS = chow sedentary; COXIV = cytochrome c oxidase subunit 4; DRP1 = dynamin-related protein 1; HE = high fat diet exercise; HFD = high fat diet; HS = high fat diet sedentary; LCLAT1 = lysocardiolipin acyltransferase 1; MFN2 = mitofusin-2; OPA1 = optic atrophy 1; PGC-1α = peroxisome proliferator-activated receptor gamma coactivator-1α; TOM70 = translocase of outer mitochondria membrane 70; VET = voluntary exercise training.

Article Snippet: The plate was centrifuged at 500 g for 5 min at 4°C, and 140 μL of MAS buffer containing 10 μg/mL cytochrome C (Thermo Fisher Scientific) was pipetted to each well.

Techniques: Expressing, Western Blot, Isolation, Marker, Control, Membrane

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: Voluntary exercise alters left ventricle gene and protein expression related to mitochondrial biogenesis and dynamics in high fat- and chow-fed mice. (A) Western blot analysis to assess purity of crude mitochondria isolation where left ventricle mitochondrial pellet expresses mitochondrial marker TOM70 and COXIV and supernatant expresses calnexin. Protein expression of (B) LCLAT1 and (C) MFN2 in left ventricle-isolated crude mitochondria and (D) LCLAT1, (E) PGC-1α, and (F) MFN2 in left ventricle tissue from male VET or sedentary mice fed an HFD or chow diet. Left ventricle mRNA expression of (G) OPA1 and (H) DRP1 in male VET or sedentary mice fed an HFD or chow diet. Values were calculated relative to 18S housekeeper. Calnexin and β-actin were used as internal controls of protein loading in left ventricle samples and COXIV as internal control of protein loading in crude mitochondria samples. 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. (B–C) n : 5–6 per group. (D–F) n = 7 per group. (G and H) n = 9 per group. CE = chow exercise; CS = chow sedentary; COXIV = cytochrome c oxidase subunit 4; DRP1 = dynamin-related protein 1; HE = high fat diet exercise; HFD = high fat diet; HS = high fat diet sedentary; LCLAT1 = lysocardiolipin acyltransferase 1; MFN2 = mitofusin-2; OPA1 = optic atrophy 1; PGC-1α = peroxisome proliferator-activated receptor gamma coactivator-1α; TOM70 = translocase of outer mitochondria membrane 70; VET = voluntary exercise training.

Article Snippet: Immunoblotting was performed with antibodies against lysocardiolipin acyltransferase 1(LCLAT1, PA5-25627; Thermo Fisher Scientific), PGC-1α (ab191838; Abcam, Cambridge, UK), MFN2 (PA5-118059; Thermo Fisher Scientific), ANP (sc-18811; Santa Cruz, Dallas, TX, USA), tumor necrosis factor alpha (TNFα, 3707; Cell Signaling, Danvers, MA, USA), phosphorylated adenosine monophosphate-activated protein kinase (AMPK, 2531; Cell Signaling), AMPKα (2532; Cell Signaling), Perilipin 5 (PA1-46215; Thermo Fisher Scientific), translocase of outer mitochondria membrane 70 (TOM70, 65675; Cell Signaling), cytochrome c oxidase subunit 4 (COXIV, 4844; Cell Signaling), and calnexin (208880; Merck, Rahway, NJ, USA).

Techniques: Expressing, Western Blot, Isolation, Marker, Control, Membrane

Journal: Journal of Radiation Research

Article Title: Mitochondrial-targeted SS-31 peptide attenuates radiation-induced cardiomyocyte senescence

doi: 10.1093/jrr/rrag048

Figure Lengend Snippet: SS-31 reduces radiation-induced mitochondrial-related apoptosis proteins in H9C2 cells and hiPSC-CMs . (A, C) Representative western blot images showing the expression of mitochondrial apoptosis-related proteins Bax and Bcl-2 in H9C2 cells (A) and hiPSC-CMs (C). (B, D) Quantification of corresponding protein expression levels, n = 3–10. Experimental groups include control, SS-31 treatment, 10 Gy radiation, and 10 Gy radiation plus SS-31 treatment, as indicated in the figure. (E) Representative western blot images of cytochrome c (Cyt c) distribution. Protein levels were analyzed in isolated mitochondrial and cytosolic fractions. (F) Quantitative analysis of Cyt c expression. Cyt c levels were normalized to VDAC1 for the mitochondrial fraction and β-actin for the cytosolic fraction. n = 3–4. All data are presented as mean ± SEM. * P < 0.05 vs control; ** P < 0.01 vs control; # P < 0.05 vs irradiation; ## P < 0.01 vs irradiation.

Article Snippet: Membranes were blocked with 1% BSA or 1% non-fat milk in tris-buffered saline with Tween 20 (TBST) for 1 hour at room temperature, followed by overnight incubation at 4°C with the following primary antibodies: TNFα (MILLIPORE, AB1837P, 1:2000), IL-6 (Sigma, SAB5700632, 1:2000), p16 (Invitrogen, PA5–20379, 1:2000), p21 (Invitrogen, 14–6715-81, 1:2000), Bax (Invitrogen MA5–14003, 1:1000), Bcl-2 (Abcam, ab196495, 1:2000), Lamin B1 (CST, 13435, 1:2000), Caspase 7 (CST, 9492S, 1:2000), Caspase 8 (CST, 4790S, 1:2000), Caspase 9 (CST, 9508S, 1:1000), Cytochrome c (Medchemexpress, HY- P80102 , 1:3000), VDAC1 (Medchemexpress, HY- P80369 , 1:3000), OxPhos Rodent WB Antibody Cocktail (Invitrogen, 45–8099, 1:2000), or β-actin (Bio-techne, MAB8929, 1:5000).

Techniques: Western Blot, Expressing, Control, Isolation, Irradiation