Journal: Mediators of Inflammation

Article Title: Enriched Environment-Induced Neuroprotection against Cerebral Ischemia-Reperfusion Injury Might Be Mediated via Enhancing Autophagy Flux and Mitophagy Flux

doi: 10.1155/2022/2396487

Figure Lengend Snippet: (a) The results of Drp1+Tomm20 costaining. (b) Comparison of the proportion of Drp1-positive mitochondria. (c) The results of Parkin+Tomm20 costaining. (d) Comparison of the proportion of Parkin-positive mitochondria. ∗∗ P < 0.01 and ∗∗∗ P < 0.001 vs . sham+SC group; # P < 0.05 and ## P < 0.01 vs . tMCAO+SC group.

Article Snippet: The sections were then sealed with 10% goat serum for 1 h and then incubated with Tomm20 antibody (H00009804-M01, Abnova).

Techniques: Comparison

Journal: Metabolism: clinical and experimental

Article Title: Krüppel-like Factor 10 Protects Against Metabolic Dysfunction-Associated Steatohepatitis By Regulating HNF4α-mediated Metabolic Pathways

doi: 10.1016/j.metabol.2024.155909

Figure Lengend Snippet: (A) Western blot assays of proteins isolated from MASH (top panel) or MAFLD (bottom panel) patients and normal individuals (n=6 per group). (B-C) Hepatic KLF10 protein levels in mice fed a high-fat/cholesterol/fructose (HFCF) diet (B, top panel), Western diet (WD) (B, bottom panel) or high-fat diet (HFD) (C, top panel) for 16 weeks (n=6 per group), or in db/db or ob/ob mice (C, bottom panel) (n=4 per group). CD, chow diet. B6, C57BL/6J mice. (D) Quantitative protein levels for studies described in A-C. (E-F) Primary hepatocytes were treated with bovine serum albumin (BSA; Veh), 100 μM palmitic acid (PA), oleic acid (OA), or linoleic acid (LA) or 10 μg/ml cholesterol (CHO) for 48 hours. Western blot assays were performed (E) and protein levels were quantified (F) (n=4 per group). Data are expressed as mean±SEM. Statistical analysis was performed using a 2-tailed, unpaired t-test (D) or one-way ANOVA (D (db/db or ob/ob mice), F). *P<0.05, **P<0.01

Article Snippet: The antibody against KLF10 (cat # H00007071-M15) was purchased from Abnova.

Techniques: Western Blot, Isolation

Journal: Metabolism: clinical and experimental

Article Title: Krüppel-like Factor 10 Protects Against Metabolic Dysfunction-Associated Steatohepatitis By Regulating HNF4α-mediated Metabolic Pathways

doi: 10.1016/j.metabol.2024.155909

Figure Lengend Snippet: (A-C) Transient transfections were performed by co-transfecting CMV-Null or CMV-KLF10 with pGL3-HNF4α (−1.0 kb WT) (A), pGL3- HNF4α (−0.3 kb WT) (B) or pGL3- HNF4α (−0.3 kb Mutant) (C) into HepG2 cells (n=7). After 24 h, relative luciferase activities (RLU) were determined after normalizing to beta-galactosidase activity. WT, wild-type. (D) C57BL/6 mice were i.v. injected with AAV8-ALB-Null or AAV8-ALB-hKLF10, then fed a Western diet for 16 weeks. ChIP assays were performed using liver lysates overexpressing KLF10 and IgG (control) or the KLF10 antibody (n=3). The DNA enrichment in the Hnf4α promoter was determined by qRT-PCR using primers that amplified a fragment between −227 bp and −154 bp (D). Statistical analysis was performed using a 2-tailed, unpaired t-test (A-D). **P<0.01

Article Snippet: The antibody against KLF10 (cat # H00007071-M15) was purchased from Abnova.

Techniques: Transfection, Mutagenesis, Luciferase, Activity Assay, Injection, Western Blot, Control, Quantitative RT-PCR, Amplification

Journal: Animals : an Open Access Journal from MDPI

Article Title: Dietary Supplementation of 25-Hydroxycholecalciferol Improves Livability in Broiler Breeder Hens-Amelioration of Cardiac Pathogenesis and Hepatopathology

doi: 10.3390/ani9100770

Figure Lengend Snippet: Effect of dietary 25-OH-D3 supplementation on cardiac hypertrophic remodeling in broiler breeder hens provided with restricted or ad libitum feed intake. At age of 29, 35, and 47 weeks, 4, 7, and 7 hens from each group were randomly selected for tissue collection. Of the hearts collected at 35 and 47 weeks, 4 hearts from each group were used for calcineurin activity analysis (panel A ), and NFATc4 activation and MHC-β expression (panels B , C ) through Western blot study. The remaining 3 hearts were used for histological studies. Results of Western blot studies were normalized to H2A.X or β-actin. All results are expressed as ratios relative to R- hens at 29 weeks. *; significant difference by ad libitum feeding (vs. corresponding R hens, p < 0.05). +; significant difference by 25-OH-D3 inclusion (vs. R or Ad hens, p < 0.05). R; restriction, Ad; ad libitum, 25-OH-D3; 25-hydroxycholecalciferol, H2A.X; H2A histone family member X, NFATc4; nuclear factor of activated T-cells, cytoplasmic 4, MHC-β; Myosin heavy chain, cardiac muscle beta, or MHC7.

Article Snippet: Heart ventricle homogenates in RIPA buffer and nuclear extracts by commercial kits (ab113474, Abcam, Cambridge, UK) were used for Western blot analysis using specific antibody against chicken MHC-β (MHC-β; Myosin heavy chain, cardiac muscle beta, clone 2E9, Developmental Studies Hybridoma Bank, Iowa City, IA, USA), and antibodies cross-reactive to chicken antigens, including rabbit anti-β-actin (Cat. # 4967, Cell Signaling Technology, Danvers, MA, USA), anti-H2A.x (H2A histone family member X, Cat. # PAB8764, Abnova Corporation, Taipei, Taiwan), anti-HIF-1α (hypoxia-inducible factor 1 alpha, Cat.# NB100-449, Novus Biologicals, Littleton, CO, USA), anti-NFATc4 (nuclear factor of activated T-cells, cytoplasmic 4, Cat. # sc-13036).

Techniques: Activity Assay, Activation Assay, Expressing, Western Blot