Fig 1: Hepatic CYP2E1 and glutathione levels and serum and molecular markers of liver injury in mice with hepatocyte-specific Il11 deletion and control mice. (A) Representative immunohistochemistry images (scale bars, 200 µm; representative dataset from n = 3 mice/group). (B) Schematic of induction of APAP injury in wild-type (WT) and CKOIl11 mice: overnight-fasted mice were intraperitoneally injected with saline or APAP (400 mg kg−1); livers and serum were collected either 0.5, 6, or 24 h following APAP administration for experiments shown in (C–F) and Figure 6. (C) Hepatic GSH concentrations at 0.5 and 6 h post-APAP. (D) ALT, (E) AST, and (F) Western blots of IL11, NOX4, GAPDH, p-ERK, ERK, p-JNK, JNK, Cl. CASP3, and CASP3 at 6 h post-APAP. (G) Representative H&E-stained liver images (scale bars, 200 µm) at 6 h post-APAP dosing quantification of the liver necrotic area. (C–E,G) Data are shown in a box-and-whisker plot with median (middle line), 25th–75th percentiles (box), and min-max values (whiskers), with a 2-way ANOVA with Sidak’s correction; 0.5 h time point: saline (n = 4 mice/genotype), APAP (n = 4 mice/genotype), 6 h time point: saline (n = 5 mice/genotype), APAP (n = 6 mice/genotype).
Fig 2: Activation of de novo ceramide synthesis in MASH models.(A) Representative H&E, Oil Red O (ORO) and Masson Trichrome (MT) staining of mouse liver tissue from the MCD diet–induced MASH model. Scale bar, 100 μm. (B) Serum ALT levels. (C) Serum AST levels. (D) Hepatic Tnf-α mRNA expression. (E) Hepatic Il-1β mRNA expression. (F) Hepatic total ceramide levels in MASH mice. (G) Heatmap of hepatic ceramide species, presented as fold change to the healthy group. Relative concentrations of each detected ceramide species were provided in data S1. (H and I) Hepatic Sptlc2 mRNA and SPTLC2 protein expression across different stages of MCD diet–induced MASH. (J and K) Hepatic Sptlc2 mRNA expression in the CD-HFD (24 weeks on diet) and STAM (8 weeks old) mouse models. N = 3 mice per group. *P < 0.05, **P < 0.01, and ***P < 0.001; [(B) to (E) and (H)] two-way ANOVA; [(F), (G), and (I)] one-way ANOVA; [(J) and (K)] unpaired Student’s t test.
Fig 3: Therapeutic efficacy of LNP-SPTLC2 siRNA in CD-HFD diet–induced MASH.(A) Schematic illustration of the experimental design. Control treatment, LNP-Scrambled siRNA; siRNA treatment, LNP-SPTLC2 siRNA. Dosing regimen: 0.3 mg siRNA/kg, administered twice weekly for 4 weeks. Created in BioRender. C. Huang (2025). https://BioRender.com/favp6kf. (B) Hepatic levels of total sphingomyelins and ceramides in MASH mice after treatment. (C) Species of main ceramides in the MASH mouse liver after treatment, the relative concentrations of each detected ceramide species are included in data S1. (D) Liver histology analysis by H&E staining (scale bar, 200 μm) for steatosis and inflammation and ORO staining (scale bar, 500 μm) for lipid content. (E) Quantification of lipid content by ORO staining. (F) Liver markers for inflammation (Il-1β, Tnf-α, and Ccl2). (G) Serum AST and AST levels. Healthy mice, n = 5; Untreated MASH mice, n = 4; control and siRNA treatment groups, n = 7 mice per group. *P < 0.05, **P < 0.01, and ***P < 0.001; one-way ANOVA.
Fig 4: Bioaccumulation of gold nanoparticles did not induce toxicities. A) Single dose accumulation. Mice (n = 5) were intravenously or intraperitoneally injected with a single dose (300 µg) of GNPs (acute dosing). After 24 h, liver, spleen, kidneys, lungs, heart, brain, ovary, and pancreas were processed to quantify GNP content using INAA. Accumulation is shown as µg GNPs/mg dry tissue. B–D) Multiple dose accumulation. Mice were intravenously or intraperitoneally injected with various doses (100, 200, and 300 µg) of GNPs every other day for 14 days (chronic dosing). The liver, spleen, kidneys, lungs, heart, brain, ovary, and pancreas were processed to quantify GNP content using INAA. Accumulation is shown as µg GNPs/mg dry tissue. E,F) Plasma were collected from mice receiving single doses (acute dosing) and ALT and AST were measured by colorimetric analysis as a measure of liver toxicity. G) Body weights of mice receiving single doses were assessed twice in 24 h. H,I) Plasma was collected from mice receiving multiple doses (chronic dosing) and ALT and AST were measured by colorimetric analysis as a measure of liver toxicity. J) The body weights of mice receiving multiple doses were assessed for 14 days.
Fig 5: HFD‐MetR changes hepatic mass and hepatokine secretion without causing injury. Livers from HFD and HFD‐MetR mice were collected, weighed, and processed for downstream assays. (a) Total alanine aminotransferase (AST) activity, (b) AST activity normalized to body weight, (c) Total aspartate aminotransferase (ALT) activity, (d) ALT activity normalized to body weight, and (e) AST to ALT ratio. (f) Total RNA from liver was isolated and reverse transcribed to quantify relative gene expression of fibrosis markers Acta2, Actin alpha2; Col1a1, collagen type 1 alpha1; Col1a2, Collagen type 1 alpha2. Quantification of (g) liver mass as percentage of total body weight. Hepatokines and enzymes were measured in blood plasma collected at the end of the experiment. Shown are quantification of (h) insulin‐like growth factor 1 (IGF‐1) and (i) Fibroblast growth factor 21 (FGF21). (j) Relative gene expression [ΔΔCq(fold change) – Relative to Actin] of insulin sensitivity markers in liver tissue; Atgl, Adipose triglyceride lipase; Scd1, stearoyl‐CoA desaturase 1; Cd36, Cluster of differentiation 36. Values shown are mean ± StdDev, with significant p values indicated. (k) Enrichment of metabolites in various metabolic pathways are shown in each group. (l) Using one‐way ANOVA comparing all profiled metabolites, 15 were identified as statistically different between groups, with p < 0.05 and log2(fold change) greater than 1.0 compared to the HFD‐control group. Abundances are these metabolites are shown in a heatmap. Each column is a sample, and each row is a metabolite. (m) The dot plot displays the enriched pathways, with significance levels on the x‐axis. The significance threshold (p = 0.05) is the gray dashed line. The size of the dots reflects the number of metabolites statistically different between groups. The color represents the enrichment, which indicates the extent to which the metabolites in the pathway were clustered amongst the compounds with the most positive fold‐changes (red), the most negative fold‐changes (blue), or uniformly distributed across high and low fold‐change compounds (white). (n) Heatmaps showing metabolite levels in the beta‐alanine metabolism pathway and (o) arginine and proline metabolism pathways.
Supplier Page from Abcam for Aspartate Aminotransferase Activity Assay Kit