Journal: bioRxiv

Article Title: Pharmacological inhibition of G protein-coupled receptor kinase 5 decreases high-fat diet-induced hepatic steatosis in mice

doi: 10.1101/2025.06.25.661655

Figure Lengend Snippet: (A) Study design showing weeks of age and treatment. Six-week-old male C57Bl/6J mice were fed a high fat diet (HFD; 45% fat, D12451, Research Diets Inc) for 8 weeks to induce obesity prior to starting water control (open square), 25 mg/kg (closed square) or 50 mg/kg (closed circle) GRK5-IN-2 treatment. After 9 weeks of treatment, metabolic phenotyping started including EchoMRI, intraperitoneal insulin and glucose tolerance tests (IPITT and IPGTT), TSE PhenoMaster chambers, and a lipogenesis functional study. Mice were euthanized after 16 weeks of treatment (Sac). (B) Body weight was measured weekly (n=10/group). (C) Body composition was measured after 9 weeks of treatment (n=10/group). (D-E) After 11-12 weeks of treatment, mice (n=4/group) were used for indirect calorimetry (TSE PhenoMaster System). Results analyzed using the CalR Web Application (Version 1.3). Gray shading indicates a dark cycle. (F-G) Mice (n=10/group) were fasted for 16 and 4 hours in preparation for an IPGTT and IPITT, respectively. Blood glucose was measured at 0, 15, 30, 60, 90, and 120-minutes post injection. Area under the curve (AUC) was calculated to assess glucose and insulin sensitivity. All results are mean ± SEM. Statistical significance was assessed using one-way or two-way ANOVA.

Article Snippet: After 11-12 weeks of treatment, mice spent 5 days single-housed in the TSE PhenoMaster environmental chambers (TSE Systems, Chesterfield, MO) to measure energy expenditure using indirect calorimetry where oxygen consumption, carbon dioxide production, respiratory exchange ratio (RER), food intake, energy expenditure, and locomotor activity were measured.

Techniques: Control, Functional Assay, Injection

Journal: bioRxiv

Article Title: Pharmacological inhibition of G protein-coupled receptor kinase 5 decreases high-fat diet-induced hepatic steatosis in mice

doi: 10.1101/2025.06.25.661655

Figure Lengend Snippet: (A) Study design illustrating timeline and treatments. Six-week-old male C57Bl/6J mice were fed a high-fat diet (HFD; 45% fat, D12451, Research Diets Inc.) for 8 weeks to induce obesity before initiating GRK5-IN-2 treatment. After 9 weeks of treatment, metabolic phenotyping was conducted, including EchoMRI, intraperitoneal insulin (IPITT) and glucose tolerance tests (IPGTT), TSE PhenoMaster metabolic chambers, and a lipogenesis functional assay using radiolabeled tracer. Mice were euthanized after 13 weeks of treatment (Sac). (B) Body weight measured weekly (n=10/group). (C) Body composition assessed via EchoMRI after 9 weeks of treatment (n=10/group). (D) Gonadal white adipose tissue (gWAT) was collected, fixed in 10% formalin, and stained with hematoxylin and eosin (H&E) for histological analysis (n=10/group). Representative H&E-stained gWAT images from water- and GRK5-IN-2-treated mice. (E) Triglyceride (TG) content in gWAT lipids extracted and quantified using a colorimetric assay (n=10/group). (F) RNA was extracted from gWAT (n=10/group), reverse-transcribed to cDNA, and analyzed via real-time PCR to quantify adipogenic and inflammatory gene expression normalized to 18S rRNA (endogenous control). Results are expressed as fold change relative to the water-treated group. All data are presented as mean ± SEM. Statistical significance was assessed using two-tailed Student’s unpaired t test.

Article Snippet: After 11-12 weeks of treatment, mice spent 5 days single-housed in the TSE PhenoMaster environmental chambers (TSE Systems, Chesterfield, MO) to measure energy expenditure using indirect calorimetry where oxygen consumption, carbon dioxide production, respiratory exchange ratio (RER), food intake, energy expenditure, and locomotor activity were measured.

Techniques: Functional Assay, Staining, Colorimetric Assay, Reverse Transcription, Real-time Polymerase Chain Reaction, Gene Expression, Control, Two Tailed Test