Journal: bioRxiv

Article Title: Glucokinase activity suppresses hepatic cholesterol synthesis and triglyceride accumulation: A new model for the effects of the GKRP P466L common human variant

doi: 10.64898/2026.04.07.717049

Figure Lengend Snippet: A) Western blot analysis of GKRP and GCK in livers from L-GKRPKO mice expressing either human GKRP (hGKRP) or the P446L variant. Samples were collected from mice after an overnight fast followed by 4 hours of refeeding after 13 weeks of HFD feeding. HSP90 was used as a loading control. B-D) Body weight (B), percent fat mass (C), and liver mass (% body weight) of hGKRP- and P446L-expressing mice over 13 weeks of HFD feeding (n = 8 per group). E) Glucose tolerance test on hGKRP- and P446L-expressing mice after 9 weeks on HFD diet (n = 8 per group). F) Plasma insulin levels in mice from (E) at baseline (fasted) and 15 minutes after glucose injection. Samples that measured below the assay’s limit of detection (0.1 ng/mL) were set to 0.1 (marked with a dashed line on the graph). G-L) L-GKRPKO mice expressing hGKRP and hGKRP P446L were fed a HFD for 13 weeks (n = 8 for both groups) G) Hepatic glycogen content after an overnight fast and refeeding for 4 hours. H) Hepatic TAG after overnight fast and refeeding for 4 hours. I) Plasma cholesterol after an overnight fast. J) Plasma cholesterol after an overnight fast and refeeding for 4 hours. K) Plasma TAG from overnight fasted mice. L) Plasma TAG after overnight fast and refeeding for 4 hours. Data are represented as mean ± SEM. Statistical significance was determined using either 2-way ANOVA with either Šidák post hoc test (B, C, and E) or Fisher’s Least Significant Difference (F) or Student’s t test (D, G-L). * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: For the triglyceride assay, 20 μL of 1% sodium deoxycholate was added to each well and the plate was incubated at 37 °C for 10 min. 200 μL of Infinity Triglyceride Reagent (Thermo Scientific TR22421) or Infinity Total Cholesterol Reagent (Thermo Fisher Scientific TR13421) was added and the plates were incubated at 37 °C for 10 min. Absorbance was measured at 500 nm and concentration was calculated via standard curve.

Techniques: Western Blot, Expressing, Variant Assay, Control, Clinical Proteomics, Injection

Journal: bioRxiv

Article Title: Glucokinase activity suppresses hepatic cholesterol synthesis and triglyceride accumulation: A new model for the effects of the GKRP P466L common human variant

doi: 10.64898/2026.04.07.717049

Figure Lengend Snippet: A) Volcano plot showing differentially expressed genes identified from RNA-sequencing analysis of livers from control and L-GCKKO mice fed a HCD for 1 week. Samples were obtained from mice that were fasted overnight followed by refeeding for 6 hours. (n = 3/4 control/KO). Red labels: genes upregulated in KO vs. control (FC ≥ 2, adjusted p value < 0.05, total = 44). Green labels: upregulated cholesterol synthesis genes (total = 16). Blue labels: genes downregulated in KO vs. control (FC ≤ −2, adjusted p value < 0.05, total = 15). B) Gene ontology analysis of differentially regulated genes. Labels in each bar represent the number of differentially regulated genes. C) mRNA levels of cholesterogenic genes in livers from control and L-GCKKO mice fed a HCD for 1 week (n = 9/8 control/KO). D-F) Control and L-GCKKO mice were fed a HCD for 1 week. Samples were obtained from mice that were fasted overnight followed by refeeding for 24 hours. (n=10/8 control/KO). D) Total hepatic cholesterol. E) D 2 O-labeled hepatic cholesterol. F) % of labeled cholesterol (of total cholesterol). Data are represented as mean ± SEM. Statistical significance was determined using Student’s t test ( C-F ). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: For the triglyceride assay, 20 μL of 1% sodium deoxycholate was added to each well and the plate was incubated at 37 °C for 10 min. 200 μL of Infinity Triglyceride Reagent (Thermo Scientific TR22421) or Infinity Total Cholesterol Reagent (Thermo Fisher Scientific TR13421) was added and the plates were incubated at 37 °C for 10 min. Absorbance was measured at 500 nm and concentration was calculated via standard curve.

Techniques: RNA Sequencing, Control, Labeling

Journal: bioRxiv

Article Title: Glucokinase activity suppresses hepatic cholesterol synthesis and triglyceride accumulation: A new model for the effects of the GKRP P466L common human variant

doi: 10.64898/2026.04.07.717049

Figure Lengend Snippet: A) Western blot analysis of GCK and HKII in livers from control, L-GCKKO, and HKII-overexpressing L-GCKKO mice after 1 week on HCD. Samples were obtained from mice that were fasted overnight followed by refeeding for 24 hours. HSP90 was used as a loading control. B) Glucose tolerance test on control, L-GCKKO, and HKII-overexpressing L-GCKKO mice after 3 days on HCD (n = 9 per group). C) Plasma insulin levels from mice in (B) at baseline (fasting) and 15 minutes after glucose infusion (n = 9 for all groups). Samples that measured below the assay’s limit of detection (0.1 ng/mL) were set to 0.1 (marked with a dashed line on the graph). D-I) Control, L-GCKKO, and HKII-overexpressing L-GCKKO mice fed a HCD for 1 week. Measurements were performed on mice that were fasted overnight, followed by refeeding for 24 hours. D) Hepatic glycogen content. E) Liver mRNA levels of Mlxipl and Pklr . F) Liver mRNA levels of cholesterogenic genes. (D-F control n = 10, L-GckKO n = 9, +HKII n = 10) G) Total hepatic cholesterol. H) D 2 O-labeled hepatic cholesterol. I) % of labeled cholesterol (of total cholesterol). (G-I control n = 7, L-GckKO n = 8, +HKII n = 8). Data are represented as mean ± SEM. Statistical significance was determined using either 2-way ANOVA with Tukey’s post hoc test (B-C) or 1-way ANOVA with Tukey’s post hoc test (D-I). * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.

Article Snippet: For the triglyceride assay, 20 μL of 1% sodium deoxycholate was added to each well and the plate was incubated at 37 °C for 10 min. 200 μL of Infinity Triglyceride Reagent (Thermo Scientific TR22421) or Infinity Total Cholesterol Reagent (Thermo Fisher Scientific TR13421) was added and the plates were incubated at 37 °C for 10 min. Absorbance was measured at 500 nm and concentration was calculated via standard curve.

Techniques: Western Blot, Control, Clinical Proteomics, Labeling

Journal: bioRxiv

Article Title: Targeting Microbial Bile Salt Hydrolase Reprograms Bile Acid Metabolism and Ameliorates Metabolic Dysfunction–Associated Steatohepatitis in Mice

doi: 10.64898/2026.05.12.724693

Figure Lengend Snippet: a , Liver weight. b , Liver-to-body weight ratio. c , Total hepatic triglyceride content. d , Total hepatic cholesterol content. e , Quantification of Oil Red O–positive area. f , Plasma alanine aminotransferase (ALT) levels. g , Plasma lipopolysaccharide (LPS) levels. h , Hepatic hydroxyproline content. i , Quantification of Sirius Red–positive area. j , Representative histological images of liver sections from chow diet (CD), western diet (WD), early-intervention low-dose (EI-Low), early-intervention high-dose (EI-High), late-intervention low-dose (LI-Low), and late-intervention high-dose (LI-High) groups. Top row: H&E staining; middle row: Oil Red O staining; bottom row: Sirius Red staining. Scale bars, 50 μm. Data are shown as mean ± SD. Each symbol represents one mouse. Statistical significance was determined by one-way ANOVA with Bonferroni’s multiple-comparisons test. * P < 0.05; ** P < 0.01; *** P < 0.001; **** P < 0.0001.

Article Snippet: Hepatic triglyceride and cholesterol concentrations were quantified using Triglyceride Liquid Reagents and Cholesterol Liquid Reagents kits, respectively (Pointe Scientific).

Techniques: Clinical Proteomics, Western Blot, Staining