Journal: Antioxidants

Article Title: Striking Cardioprotective Effects of an Adiponectin Receptor Agonist in an Aged Mouse Model of Duchenne Muscular Dystrophy

doi: 10.3390/antiox13121551

Figure Lengend Snippet: Effects of AdipoRon treatment on cardiac muscle fibrosis and hypertrophy. ( A ) Picro-Sirius Red staining. Scale bar = 200 µm. ( B ) Quantification of Picro-Sirius Red staining. mRNA levels of ( C ) αSMA and ( D ) TGF-β1, two markers of fibrosis. ELISA assays were used to quantify ( E ) TGF-β and ( F ) the active phosphorylated form of SMAD2 (P¬SMAD2), a transcription factor mainly involved in TGF-β signalling. mRNA levels of ( G ) BNP and ( H ) ANP, two markers of hypertrophy. ( I ) ELISA assay was also used to quantify the levels of ANP. The percentage of stained areas was calculated in cardiac muscle sections, and the subsequent ratios are presented as relative expressions to WT values. mRNA levels were normalised to cyclophilin, and the subsequent ratios were presented as relative expressions to WT values. Absorbance data are presented as relative expressions to WT values. Data are means ± SD; n = 6 mice per group for all experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. WT mice. ### p < 0.001, #### p < 0.0001 vs. mdx mice.

Article Snippet: ELISA assays were also used to quantify the levels of ANP, 4-Hydroxynonenal (HNE), TNFα, IL-1β, IL-10 (all from Abcam, Cambridge, UK), utrophin A (UTRN) (from Antibodies Online, Atlanta, GA, USA), TGF-β, AdipoR1, AdipoR2, the active phosphorylated form of Calcium/Calmodulin Dependent Protein Kinase Kinase 2 (CAMKK2), peroxisome proliferator-activated receptor α (PPARα), PPAR gamma coactivator 1 alpha (PGC-1α), Translocase of Outer Mitochondrial Membrane 20 (TOMM20), and the active phosphorylated form of Ribosome-inactivating protein (P-RIP) (all from MyBiosource—Bio-Connect Diagnostics B.V., Huissen, The Netherlands).

Techniques: Staining, Enzyme-linked Immunosorbent Assay

Journal: Antioxidants

Article Title: Striking Cardioprotective Effects of an Adiponectin Receptor Agonist in an Aged Mouse Model of Duchenne Muscular Dystrophy

doi: 10.3390/antiox13121551

Figure Lengend Snippet: Effects of AdipoRon treatment on ApN receptors and signalling in the dystrophic cardiac muscle. mRNA levels of ( A ) AdipoR1 and ( B ) AdipoR2, adiponectin main receptors. ELISA assays were used to quantify ( C ) AdipoR1 and ( D ) AdipoR2. ( E ) The ratio of AdpoR1 over AdipoR2 mRNA levels was calculated within the cardiac muscle. ELISA assays were used to quantify ( F ) the active phosphorylated form of AMPKα (P-AMPK), ( G ) calcium/calmodulin-dependent protein kinase 2 (CAMKK2), and ( H ) peroxisome proliferator-activated receptor alpha (PPARα), ApN/AdipoRon, main signalling pathways in muscle. ( I ) mRNA levels of PGC-1α. ELISA assays were used to quantify ( J ) PGC-1α, ( K ) the active phosphorylated form of the p65 subunit of NF-κB (P-p65), a transcription factor mainly involved in inflammation, and ( L ) utrophin A (UTRN), a dystrophin analogue. mRNA levels were normalised to cyclophilin, and the subsequent ratios are presented as relative expression to WT values. Absorbance data are presented as relative expressions to WT values. Data are means ± SD; n = 6 mice per group for all experiments. Statistical analysis was performed using one-way ANOVA followed by Tukey’s test. * p < 0.05, ** p < 0.01, **** p < 0.0001 vs. WT mice. # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 vs. mdx mice.

Article Snippet: ELISA assays were also used to quantify the levels of ANP, 4-Hydroxynonenal (HNE), TNFα, IL-1β, IL-10 (all from Abcam, Cambridge, UK), utrophin A (UTRN) (from Antibodies Online, Atlanta, GA, USA), TGF-β, AdipoR1, AdipoR2, the active phosphorylated form of Calcium/Calmodulin Dependent Protein Kinase Kinase 2 (CAMKK2), peroxisome proliferator-activated receptor α (PPARα), PPAR gamma coactivator 1 alpha (PGC-1α), Translocase of Outer Mitochondrial Membrane 20 (TOMM20), and the active phosphorylated form of Ribosome-inactivating protein (P-RIP) (all from MyBiosource—Bio-Connect Diagnostics B.V., Huissen, The Netherlands).

Techniques: Enzyme-linked Immunosorbent Assay, Expressing

Journal: Antioxidants

Article Title: Striking Cardioprotective Effects of an Adiponectin Receptor Agonist in an Aged Mouse Model of Duchenne Muscular Dystrophy

doi: 10.3390/antiox13121551

Figure Lengend Snippet: Effects of AdipoRon treatment on cardiac muscle oxidative capacity, injury, and overall muscle function. mRNA levels of ( A ) ERRα and ( B ) mtTFA, two markers of mitochondrial biogenesis. ELISA assay was used to quantify ( C ) TOMM20, a marker of mitochondrial content. ( D ) Wire test where mice hanging time was recorded (s). ( E ) Fore-limb grip test and ( F ) fore- and hind-limb grip test, measuring muscle strength expressed in Gram-force relative to body weight (gf/gBW). ( G ) Treadmill running exercise, where the total distance covered on the third day was measured (m). ( H ) CK and ( I ) LDH plasma activities assessing muscle injury and expressed as IU/L. ( J ) ELISA assay was used to quantify the active phosphorylated form of RIP (P-RIP), an important regulator of cellular stress that triggers a regulated pathway for necrotic cell death called necroptosis. mRNA levels were normalised to cyclophilin, and the subsequent ratios are presented as relative expressions to WT values. Absorbance data are presented as relative expressions to WT values. Data are means ± SD; n = 6 mice per group for all ex vivo experiments. Data are means ± SD; n = 7–8 mice per group for all in vivo functional tests. Statistical analysis was performed using one-way ANOVA followed by Tukey’s test. ** p < 0.01, *** p < 0.001, **** p < 0.0001 vs. WT mice. # p < 0.05, ## p < 0.01, ### p < 0.001, #### p < 0.0001 vs. mdx mice.

Article Snippet: ELISA assays were also used to quantify the levels of ANP, 4-Hydroxynonenal (HNE), TNFα, IL-1β, IL-10 (all from Abcam, Cambridge, UK), utrophin A (UTRN) (from Antibodies Online, Atlanta, GA, USA), TGF-β, AdipoR1, AdipoR2, the active phosphorylated form of Calcium/Calmodulin Dependent Protein Kinase Kinase 2 (CAMKK2), peroxisome proliferator-activated receptor α (PPARα), PPAR gamma coactivator 1 alpha (PGC-1α), Translocase of Outer Mitochondrial Membrane 20 (TOMM20), and the active phosphorylated form of Ribosome-inactivating protein (P-RIP) (all from MyBiosource—Bio-Connect Diagnostics B.V., Huissen, The Netherlands).

Techniques: Enzyme-linked Immunosorbent Assay, Marker, Ex Vivo, In Vivo, Functional Assay