plasma cholesterol  (Thermo Fisher)

Journal: The Journal of Biological Chemistry

Article Title: Cell type-specific control of cardiometabolic disease by the dileucine motif of the LDL receptor-related protein LRP1

doi: 10.1016/j.jbc.2026.111421

Figure Lengend Snippet: Hepatosteatosis and liver gene expression in Western diet-fed Lrp1 LL Ldlr −/− (LL) and Lrp1 AA Ldlr −/− (AA) mice. Age-matched LL and AA mice were fed Western diet for 16 weeks. A , livers were obtained from 7 LL and 6 AA mice for histological examination of steatosis and measurements of triglyceride and cholesterol content. Total RNA was extracted from 6 to 7 LL and 6 to 7 AA mice for RT-PCR to quantify expression of: ( B ) cholesterol synthesis genes HMGCR, HMGCS, and SREBP2, ( C ) fatty acid oxidation genes ACOX1, CYPT1α, and PDK4, and ( D ) PPARα, PPARβ, and PPARγ. The data were evaluated for statistical significance by Student’s t test with p values as shown. E , protein levels of PPARα, PPARβ/δ, and PPARγ in livers prepared from Western diet-fed LL and AA mice were assessed by Western blot analysis of liver lysates using GAPDH as loading control. The data were normalized to the mean levels of expression in LL samples, with fold changes as indicated.

Article Snippet: Plasma obtained from animals after an overnight fast was used to quantify plasma cholesterol and triglyceride levels using Infinity colorimetric assay kits (TR22421 and TR13421, Thermo Fisher Scientific).

Techniques: Gene Expression, Western Blot, Reverse Transcription Polymerase Chain Reaction, Expressing, Control

Journal: The Journal of Biological Chemistry

Article Title: Cell type-specific control of cardiometabolic disease by the dileucine motif of the LDL receptor-related protein LRP1

doi: 10.1016/j.jbc.2026.111421

Figure Lengend Snippet: Plasma lipids and lipoproteins in Western diet-fed Lrp1 LL Ldlr −/− (LL) and Lrp1 AA Ldlr −/− mice. The LL and AA mice were fed a Western-type diet for 16 weeks. A , VLDL production was assessed in fasting animals (n = 3 per group) by measuring plasma triglyceride levels and determining changes in plasma triglyceride levels (Δtriglyceride) hourly after injection of Poloxamer 407 to inhibit lipolysis. B , plasma cholesterol and triglyceride levels were measured in 14 LL and 12 AA mice after an overnight fast. C , pooled plasma samples from fasting LL and AA mice were fractionated by FPLC for lipoprotein distribution analysis. The elution profiles were compared to elutions of standard chylomicron remnants/VLDL, IDL/LDL and HDL as indicated. D , plasma apolipoproteins were identified by Western blot analysis of plasma samples (n = 8 per group) for apoB (identifying both apoB100 and apoB48), apoE, and apoAI. Levels of apoB100, apoB48, and apoE were normalized to apoAI levels in plasma. All data were evaluated for significance by Student’s t test with p values as shown.

Article Snippet: Plasma obtained from animals after an overnight fast was used to quantify plasma cholesterol and triglyceride levels using Infinity colorimetric assay kits (TR22421 and TR13421, Thermo Fisher Scientific).

Techniques: Clinical Proteomics, Western Blot, Injection

Journal: The Journal of Biological Chemistry

Article Title: Cell type-specific control of cardiometabolic disease by the dileucine motif of the LDL receptor-related protein LRP1

doi: 10.1016/j.jbc.2026.111421

Figure Lengend Snippet: LRP1 with proximal dileucine motif mutation in bone marrow cells exacerbates atherosclerosis. Bone marrows from Lrp1 LL Ldlr −/− (LL) and Lrp1 AA Ldlr −/− (AA) mice were transplanted into Lrp1 LL Ldlr −/− recipient mice and then fed the Western type diet for 16 weeks ( A ) Plasma cholesterol levels in 8 LL and 6 AA mice, ( B ) plasma triglyceride levels in 8 LL and 6 AA mice, ( C ) atherosclerotic plaque area in the whole aorta of 7 mice per group, and ( D ) atherosclerotic lesion area in the aortic roots of 7 mice per group were analyzed. Bone marrows from LL mice were also transplanted into LL and AA recipient mice. After feeding the Western diet for 16 weeks, fasting plasma samples from 10 LL and 10 AA mice were analyzed for ( E ) cholesterol and ( F ) triglyceride levels. G , atherosclerotic lesion area in the whole aortas of LL and AA mice (n = 76 per group) and ( H ) in the aortic roots (n = 10 per group) were characterized. All data were evaluated for significance by Student’s t test with p values as shown.

Article Snippet: Plasma obtained from animals after an overnight fast was used to quantify plasma cholesterol and triglyceride levels using Infinity colorimetric assay kits (TR22421 and TR13421, Thermo Fisher Scientific).

Techniques: Mutagenesis, Western Blot, Clinical Proteomics

Journal: The Journal of Biological Chemistry

Article Title: Cell type-specific control of cardiometabolic disease by the dileucine motif of the LDL receptor-related protein LRP1

doi: 10.1016/j.jbc.2026.111421

Figure Lengend Snippet: LRP1 proximal dileucine mutation reduces mitochondrial respiration in cholesterol-loaded macrophages. Bone marrow-derived macrophages were incubated with or without acLDL prior to determination of oxygen consumption rates (OCR) ( A and B ). Oligomycin, FCCP, and rotenone/antimycin (rot/AA) were added to measure OCR and determination of basal ( C ) and maximal respiration ( D ). Etomoxir was added at 3 μM to inhibit fatty acid oxidation to determine OCR for assessment of basal respiration ( E ) and maximal respiration ( F ) due to non-fatty acid sources. Total mitochondrial ATP production ( G ) and etomoxir-resistant ATP production ( H ) were calculated from the OCR data. Differences between LL and AA samples under various treatment conditions were analyzed for significance by Student’s t test. ∗ indicates p < 0.05; ∗∗ indicates p < 0.01; ∗∗∗ p < 0.001; n.s. indicates not significant.

Article Snippet: Plasma obtained from animals after an overnight fast was used to quantify plasma cholesterol and triglyceride levels using Infinity colorimetric assay kits (TR22421 and TR13421, Thermo Fisher Scientific).

Techniques: Mutagenesis, Derivative Assay, Incubation

Journal: The Journal of Biological Chemistry

Article Title: Cell type-specific control of cardiometabolic disease by the dileucine motif of the LDL receptor-related protein LRP1

doi: 10.1016/j.jbc.2026.111421

Figure Lengend Snippet: LRP1 proximal dileucine mutation increased levels of mitochondrial fission proteins in cholesterol-loaded macrophages. Bone marrow macrophages isolated from Lrp1 LL Ldlr −/− (LL) and Lrp1 AA Ldlr −/− (AA) mice (n = 2 per group) were treated with acLDL for 24 h for cholesterol loading. Protein lysates were prepared for Western blot analysis of mitochondrial fission proteins: ( A ) mitochondrial fission factor MFF and ( B ) dynamin-related protein-1 DRP1; as well as mitochondrial fusion proteins: ( C ) mitofusin-2 (MFN2) and ( D ) optic atrophy-1 (OPA1). β-actin level in each sample was used as loading control. The data were normalized to the mean levels of expression in LL samples, with fold changes as indicated.

Article Snippet: Plasma obtained from animals after an overnight fast was used to quantify plasma cholesterol and triglyceride levels using Infinity colorimetric assay kits (TR22421 and TR13421, Thermo Fisher Scientific).

Techniques: Mutagenesis, Isolation, Western Blot, Control, Expressing

Journal: The Journal of Biological Chemistry

Article Title: Cell type-specific control of cardiometabolic disease by the dileucine motif of the LDL receptor-related protein LRP1

doi: 10.1016/j.jbc.2026.111421

Figure Lengend Snippet: LRP1 proximal dileucine motif mutation does not impact aggLDL uptake by reduced LRP1 association with mitochondria in cholesterol-loaded macrophages. A , bone marrow macrophages isolated from Lrp1 LL Ldlr −/− (LL) and Lrp1 AA Ldlr −/− mice (n = 6 per group) were incubated with DiI-labeled aggLDL overnight. Intracellular DiI accumulation was determined by fluorometry to determine cellular uptake of aggLDL. B , the bone marrow macrophages were treated with acLDL for 24 h for cholesterol loading. Total cell lysates and mitochondria fractions were prepared for Western blot analysis of total LRP1 levels and LRP1 association with mitochondria. β-actin levels in cell lysates and ATP5A levels in mitochondrial fractions were used as loading control. The data were normalized to the mean levels of expression in LL samples, with fold changes as indicated.

Article Snippet: Plasma obtained from animals after an overnight fast was used to quantify plasma cholesterol and triglyceride levels using Infinity colorimetric assay kits (TR22421 and TR13421, Thermo Fisher Scientific).

Techniques: Mutagenesis, Isolation, Incubation, Labeling, Western Blot, Control, Expressing

Journal: bioRxiv

Article Title: Targeted Epigenetic Modulation Outperforms Nuclease- and Deaminase-Based Editing for Durable Pcsk9 Silencing in a Clinically Relevant Delivery System

doi: 10.64898/2026.03.20.713290

Figure Lengend Snippet: (A) Experimental workflow. Hepa1-6 mRNA (epi-editor, gene editor, or base editor) and sgRNA were formulated LUNAR ® for systemic delivery, while GalNAc-conjugated siRNA was administered directly. eGFP mRNA served as a vehicle control. Blood samples were collected at the indicated time points for downstream analyses. Created with BioRender.com. (B–C) Serum aspartate aminotransferase (AST) and alanine aminotransferase (ALT) levels measured at each time point to assess hepatotoxicity. (D) Plasma PCSK9 protein levels quantified by ELISA. (E–F) Circulating low-density lipoprotein cholesterol (LDL-C) and total cholesterol levels. All plasma data (D–F) are normalized to eGFP control at the corresponding time point. Statistical analyses were performed as described in the Methods section. Error bars: SD.

Article Snippet: Plasma low-density lipoprotein cholesterol (LDL-C) was measured using a colorimetric assay kit (Novus Biologicals, Centennial, CO) following the manufacturer’s protocol, with samples loaded undiluted.

Techniques: Control, Clinical Proteomics, Enzyme-linked Immunosorbent Assay

Journal: bioRxiv

Article Title: Targeted Epigenetic Modulation Outperforms Nuclease- and Deaminase-Based Editing for Durable Pcsk9 Silencing in a Clinically Relevant Delivery System

doi: 10.64898/2026.03.20.713290

Figure Lengend Snippet: (A) Body weights of 8–12-week-old female C57BL/6N mice following intravenous administration of 1 mg/kg RNA formulated in LUNAR ® . (B–D) Plasma levels of PCSK9 protein (B), low-density lipoprotein cholesterol (LDL-C) (C), and total cholesterol (D) measured at indicated time points post-dosing. (E) Comparison of therapeutic efficacy of different gene-modifying modalities on plasma PCSK9, LDL-C, and total cholesterol levels at day 30 post-dose. Statistical significance was determined by one-way ANOVA with Tukey’s multiple comparisons test (* P < 0.05, *** P < 0.001). For PCSK9 comparisons, differences between groups not specifically indicated are all statistically significant ( P < 0.001). Error bars: SD.

Article Snippet: Plasma low-density lipoprotein cholesterol (LDL-C) was measured using a colorimetric assay kit (Novus Biologicals, Centennial, CO) following the manufacturer’s protocol, with samples loaded undiluted.

Techniques: Clinical Proteomics, Comparison, Drug discovery

Journal: bioRxiv

Article Title: Targeted Epigenetic Modulation Outperforms Nuclease- and Deaminase-Based Editing for Durable Pcsk9 Silencing in a Clinically Relevant Delivery System

doi: 10.64898/2026.03.20.713290

Figure Lengend Snippet: (A) Experimental design comparing the efficacy of epi-editor delivery between the Capelluti et al. study and Arcturus Therapeutics. Plasma was collected at the indicated time points (dots colored by time). (B) Plasma PCSK9 levels measured at each time point post-dosing. Data were normalized to the respective vehicle controls (PBS-treated mice for Capelluti et al. and LUNAR ® -formulated eGFP mRNA for Arcturus data). (C–E) Comparison of plasma PCSK9 (C), LDL-C (D), and total cholesterol (E) at day 30 post-dosing. Values were normalized to the respective vehicle controls. Error bars: SD. Statistical significance was determined by one-way ANOVA followed by Tukey’s multiple comparisons test (* P < 0.05, ** P < 0.01, *** P < 0.001). Capelluti’s data were adapted from provided data in and Arcturus data were from the current study.

Article Snippet: Plasma low-density lipoprotein cholesterol (LDL-C) was measured using a colorimetric assay kit (Novus Biologicals, Centennial, CO) following the manufacturer’s protocol, with samples loaded undiluted.

Techniques: Clinical Proteomics, Comparison

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Cysteine 129 in Pla2g12b Is Critical for Intestinal and Hepatic Lipoprotein Secretion in Mice

doi: 10.1016/j.jcmgh.2025.101712

Figure Lengend Snippet: Pla2g12b mutant mice have lower plasma lipid and lipoprotein levels than WT mice. ( A and B ) Male ( A ) and female ( B ) WT and MUT siblings (n = 4/group) were weaned onto a chow diet. Plasma was collected from overnight fasted 3-month-old mice, and total cholesterol and TG, as well as cholesterol in HDL and LDL, were measured after polyethylene glycol precipitation and correction with Friedewald’s equation. ( C and D ) Pooled plasma (250 μL) from male ( C ) and female ( D ) mice was subjected to gel filtration, and cholesterol and TG levels were measured in each fraction. ( E and F ) Plasma (1 μL) was used to detect apoB in male ( E ) and female ( F ) mice via immunoblotting ( top ). Bands were quantified and plotted ( bottom ). Data are representative of 3 independent group measurements. ns, not significant; ∗∗ P < .01; ∗∗∗ P < 01; ∗∗∗∗ P < .0001; Student’s t -test.

Article Snippet: Plasma cholesterol (Fisher Scientific, #23-666-202) and TG (Fisher Scientific, #23-666-412) levels were measured.

Techniques: Mutagenesis, Clinical Proteomics, Filtration, Western Blot

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Cysteine 129 in Pla2g12b Is Critical for Intestinal and Hepatic Lipoprotein Secretion in Mice

doi: 10.1016/j.jcmgh.2025.101712

Figure Lengend Snippet: Livers from MUT mice show greater lipid accumulation than WT mice. ( A ) Livers were collected from fasted male (n = 4 per group) and female mice (n = 5 per group) and photographed. A representative picture is shown. Livers from WT mice were reddish; in contrast, those from MUT mice were paler, indicating lipid accumulation. ( B ) Shortly after sacrifice, organs were collected, washed in cold PBS by dipping, blotted dry, and weighed. Wet weights of livers were significantly higher in MUT than WT mice. ( C ) Small liver slices from different mice were embedded and used for H&E staining ( top ) and electron microscopy ( bottom ). H&E staining and electron microscopy micrographs showed large vacuoles and large droplets in the livers in MUT mice. ( D ) Trichome staining showed no deposition of collagen in both the male WT and MUT livers. ( E ) Hepatic cholesterol and TG content was higher in MUT mice than controls. ( F ) Plasma ALT and AST were higher in plasma of MUT mice than controls. ns, not significant; ∗ P < .05; ∗∗ P < .01; ∗∗∗ P < .001; ∗∗∗∗ P < .0001; Student’s t -test.

Article Snippet: Plasma cholesterol (Fisher Scientific, #23-666-202) and TG (Fisher Scientific, #23-666-412) levels were measured.

Techniques: Staining, Electron Microscopy, Clinical Proteomics

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Cysteine 129 in Pla2g12b Is Critical for Intestinal and Hepatic Lipoprotein Secretion in Mice

doi: 10.1016/j.jcmgh.2025.101712

Figure Lengend Snippet: Pla2g12b MUT mice are defective in lipid absorption. ( A ) Overnight fasted male WT (n = 4) and MUT (n = 5) mice were gavaged with olive oil (200 μL/mouse). Plasma was collected at the indicated times, and TG levels were measured. Plasma TG levels increased over time in WT mice but not in MUT mice, thus indicating a defect in lipid absorption in MUT mice. Repeated measures mixed-effect model ANOVA, group × time. ( B ) Overnight fasted 3-month-old female mice (n = 4/group) were gavaged (200 μL olive oil/mouse), plasma was collected at various times, and TG and cholesterol levels were measured. Plasma TG excursions were severely blunted in MUT mice, thus indicating defects in fat absorption. ( C ) Overnight fasted male WT (n = 4) and MUT (n = 3) mice were gavaged with olive oil (200 μL/mouse) containing 2 mg of NBD-TG. Blood was collected at indicated times. Total plasma fluorescence ( left ) and TG ( right ) were measured. Repeated measures mixed-effect model ANOVA, group × time. ( D ) In a separate experiment, WT and MUT mice were placed in CLAMS to collect feces over a period of 5 days. After drying, 100 mg fecal powder was used for lipid extraction and quantification. ns, not significant; ∗ P < .05; Student’s t -test.

Article Snippet: Plasma cholesterol (Fisher Scientific, #23-666-202) and TG (Fisher Scientific, #23-666-412) levels were measured.

Techniques: Clinical Proteomics, Fluorescence, Extraction

Journal: Cellular and Molecular Gastroenterology and Hepatology

Article Title: Cysteine 129 in Pla2g12b Is Critical for Intestinal and Hepatic Lipoprotein Secretion in Mice

doi: 10.1016/j.jcmgh.2025.101712

Figure Lengend Snippet: Pla2g12b MUT mice accumulate lipids in intestine. ( A ) Three hours after oral gavage, duodenum and jejunum samples were sectioned and stained with H&E. Both the duodenum and jejunum in MUT mice showed large vacuoles indicative of greater lipid accumulation than observed in WT controls. ( B ) Intestinal sections were also stained with OROO. Intestines from MUT mice showed significantly more red staining than control intestines, thus indicating lipid accretion. ( C ) After 3 hours, tissues were collected and cholesterol and TG levels in the duodenum and jejunum were measured. Lipid levels were higher in the tissues in the MUT mice. ns, not significant; ∗ P < .05; ∗∗∗ P < .01; Student’s t -test and 2-way ANOVA. ( D ) Overnight fasted 3-month-old female mice (n = 4/group) were gavaged (200 μL olive oil/mouse). After 5 hours, the mice were sacrificed, duodenum and jejunum were collected, and lipid levels were measured. Tissues from MUT mice had higher cholesterol and TG levels than those from control mice. ∗ P < .05; ∗∗ P < .01; ∗∗∗∗ P < .0001; Student’s t -test.

Article Snippet: Plasma cholesterol (Fisher Scientific, #23-666-202) and TG (Fisher Scientific, #23-666-412) levels were measured.

Techniques: Staining, Control