Journal: Journal of Lipid Research

Article Title: A decreased expression of angiopoietin-like 3 is protective against atherosclerosis in apoE-deficient mice

doi: 10.1194/jlr.m300031-jlr200

Figure Lengend Snippet: Fig. 1. Angiopoietin-like 3 (Angptl3) expression in the liver of mice with a recessive mutation in Angptl3 gene causing hypolipidemia (hypl) in the presence (A) and absence (B) of apolipoprotein E (apoE). TaqMan analyses were performed in the liver of 24-week-old males (n 4) fasted for 16 h. The amount of Angptl3 mRNA was corrected by dividing it with that of 36B4 mRNA in each sample. Values are depicted as mean SD. *P 0.05, **P 0.01.

Article Snippet: Total RNA was extracted from the liver using Trizol Reagent (Invitrogen) and subjected to reverse transcription and amplification according to the protocol supplied with the TaqMan Gold RTPCR kit (Perkin-Elmer) for Angptl3 and 36B4 gene.

Techniques: Expressing, Mutagenesis

Journal: Journal of Molecular Cell Biology

Article Title: ANGPTL3 negatively regulates IL-1 β -induced NF- κ B activation by inhibiting the IL1R1-associated signaling complex assembly

doi: 10.1093/jmcb/mjad053

Figure Lengend Snippet: Overexpression of ANGPTL3 inhibits IL-1β-induced NF-κB activation. ( A and B ) Overexpression of ANGPTL3 inhibits IL-1β-induced NF-κB activation in a dose-dependent manner as determined by the reporter assay in HEK293T (upper panel in A ) and Hep3B (upper panel in B ) cells. The expression levels of ANGPTL3-Flag in the indicated cell lysate were detected by western blotting (lower panels). ( C – E ) The effects of overexpressed ANGPTL3 on IL-1β-induced transcription of TNFA, CXCL2 , and IL8 were determined by qPCR in HEK293T ( C ), Hep3B ( D ), and primary hepatocytes ( E ). AN3, ANGPTL3. ** P < 0.01; *** P < 0.001.

Article Snippet: Antibodies against ANGPTL3 (R&D Systems, AF3829), MyD88 (Cell Signaling Technology, 4283), Flag tag (Sigma, F1804), HA tag (Sigma, H3663), and Myc tag (Cell Signaling Technology, 2276) were purchased.

Techniques: Over Expression, Activation Assay, Reporter Assay, Expressing, Western Blot

Journal: Journal of Molecular Cell Biology

Article Title: ANGPTL3 negatively regulates IL-1 β -induced NF- κ B activation by inhibiting the IL1R1-associated signaling complex assembly

doi: 10.1093/jmcb/mjad053

Figure Lengend Snippet: Knockdown of ANGPTL3 potentiates IL-1β-induced NF-κB activation. ( A ) ANGPTL3 shRNA plasmids significantly reduced the protein levels of both transfected Flag-tagged ANGPTL3 (upper panel, in HEK293T cells) and endogenous ANGPTL3 (lower panel, in Hep3B cells). ( B and C ) The effects of ANGPTL3 knockdown on IL-1β-induced NF-κB activation were determined by the reporter assay in HEK293T ( B ) and Hep3B ( C ) cells. ( D – F ) The effects of ANGPTL3 knockdown on IL-1β-induced transcription of TNFA, CXCL2 , and IL8 were determined by qPCR in Hep3B ( D ), HepG2 ( E ), and HEK293T ( F ) cells. ( G and H ) The effects of ANGPTL3 knockdown on IL-1β- ( G ) or LPS-induced ( H ) transcription of TNFA, CXCL2 , and IL8 were determined by qPCR in THP1 cells. AN3, ANGPTL3; Exo., exogenous; Endo., endogenous. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant.

Article Snippet: Antibodies against ANGPTL3 (R&D Systems, AF3829), MyD88 (Cell Signaling Technology, 4283), Flag tag (Sigma, F1804), HA tag (Sigma, H3663), and Myc tag (Cell Signaling Technology, 2276) were purchased.

Techniques: Knockdown, Activation Assay, shRNA, Transfection, Reporter Assay

Journal: Journal of Molecular Cell Biology

Article Title: ANGPTL3 negatively regulates IL-1 β -induced NF- κ B activation by inhibiting the IL1R1-associated signaling complex assembly

doi: 10.1093/jmcb/mjad053

Figure Lengend Snippet: Knockout of ANGPTL3 potentiates IL-1β-induced NF-κB activation. ( A ) ANGPTL3 sgRNA plasmids significantly reduced the protein levels of both transfected Flag-tagged ANGPTL3 (upper panel, in HEK293T cells) and endogenous ANGPTL3 (lower panel, in Hep3B cells). ( B and C ) The effects of ANGPTL3 deficiency on IL-1β-induced transcription of TNFA, CXCL2 , and IL8 were determined by qPCR in Hep3B ( B ) and HepG2 ( C ) cells. ( D and E ) The effects of ANGPTL3 deficiency on IL-1β- ( D ) and LPS-induced ( E ) transcription of TNFA, CXCL2 , and IL8 were determined by qPCR in THP1 cells. AN3, ANGPTL3; KO, knockout; Exo., exogenous; Endo., endogenous. * P < 0.05; *** P < 0.001; ns, not significant.

Article Snippet: Antibodies against ANGPTL3 (R&D Systems, AF3829), MyD88 (Cell Signaling Technology, 4283), Flag tag (Sigma, F1804), HA tag (Sigma, H3663), and Myc tag (Cell Signaling Technology, 2276) were purchased.

Techniques: Knock-Out, Activation Assay, Transfection

Journal: Journal of Molecular Cell Biology

Article Title: ANGPTL3 negatively regulates IL-1 β -induced NF- κ B activation by inhibiting the IL1R1-associated signaling complex assembly

doi: 10.1093/jmcb/mjad053

Figure Lengend Snippet: ANGPTL3 targets the IL1R1-associated complex. ( A ) The effects of ANGPTL3 knockdown on IL1R1–IL1RAP- or MyD88-mediated NF-κB activation were determined by the reporter assay in HEK293T cells. ** P < 0.01; ns, not significant. ( B ) The interaction of overexpressed ANGPTL3 with IL1R1, IL1RAP, MyD88, and IKKβ was determined by Co-IP in HEK293T cells. ( C ) The interaction of endogenous ANGPTL3 with IL1R1 and IL1RAP was determined by Co-IP in HepG2 cells. ( D ) The co-localization of ANGPTL3-Cherry with IL1R1-GFP and IL1RAP-GFP in HEK293T cells. Scale bar, 10 μm. ( E ) A schematic presentation of human IL1R1, IL1RAP, and their truncation mutants. ( F and G ) The interaction of ANGPTL3 with IL1R1, IL1RAP, and their truncation mutants was determined by Co-IP in HEK293T cells. AN3, ANGPTL3; FL, full-length; Ig, immunoglobulin domain; TM, transmembrane domain; TIR: Toll/IL-1R homology domain.

Article Snippet: Antibodies against ANGPTL3 (R&D Systems, AF3829), MyD88 (Cell Signaling Technology, 4283), Flag tag (Sigma, F1804), HA tag (Sigma, H3663), and Myc tag (Cell Signaling Technology, 2276) were purchased.

Techniques: Knockdown, Activation Assay, Reporter Assay, Co-Immunoprecipitation Assay

Journal: Journal of Molecular Cell Biology

Article Title: ANGPTL3 negatively regulates IL-1 β -induced NF- κ B activation by inhibiting the IL1R1-associated signaling complex assembly

doi: 10.1093/jmcb/mjad053

Figure Lengend Snippet: The intracellular FLD domain of ANGPTL3 is involved in the negative regulation of IL-1β-induced signaling. ( A ) The effects of intracellular and secreted ANGPTL3 on IL-1β-induced NF-κB activation were determined by the reporter assay in HEK293T cells. ( B ) The effects of secreted full-length or truncated ANGPTL3 on IL-1β-induced NF-κB activation were determined by the reporter assay and western blotting in HEK293T cells. ( C ) The effects of wild-type or secretion-defective mutants of ANGPTL3 on the regulation of IL-1β-induced NF-κB activation were determined by the reporter assay and western blotting in HEK293T cells. ( D ) A schematic presentation of human ANGPTL3 and its truncation mutants. ( E ) The effects of full-length and various mutants of ANGPTL3 on IL-1β-induced NF-κB activation were determined by the reporter assay in HEK293T cells. ( F ) The interaction of IL1R1 with full-length and truncation mutants of ANGPTL3 was determined by Co-IP in HEK293T cells. AN3, ANGPTL3; Sup., supernatant; FL, full-length; WT, wild-type; SP, signal peptide; CCD, coiled-coil domain; FLD, fibrinogen-like domain. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant.

Article Snippet: Antibodies against ANGPTL3 (R&D Systems, AF3829), MyD88 (Cell Signaling Technology, 4283), Flag tag (Sigma, F1804), HA tag (Sigma, H3663), and Myc tag (Cell Signaling Technology, 2276) were purchased.

Techniques: Activation Assay, Reporter Assay, Western Blot, Co-Immunoprecipitation Assay

Journal: Journal of Molecular Cell Biology

Article Title: ANGPTL3 negatively regulates IL-1 β -induced NF- κ B activation by inhibiting the IL1R1-associated signaling complex assembly

doi: 10.1093/jmcb/mjad053

Figure Lengend Snippet: ANGPTL3 inhibits the formation of IL1R1-associated complex. ( A and B ) The effects of ANGPTL3 overexpression on IL1R1 or IL1RAP homo- or hetero-interactions were determined by Co-IP ( A ) and quantitative analysis ( B ) in HEK293T cells. ( C – F ) The effects of ANGPTL3 overexpression on the IL1R1–MyD88 and IL1R1–IL-1β interactions were determined by Co-IP ( C and E ) and quantitative analysis ( D and F ) in HEK293T cells. ( G and H ) The effects of ANGPTL3 deficiency on the IL1R1–IL1RAP and IL1RAP–IL1RAP interactions were determined by Co-IP ( G ) and quantitative analysis ( H ) in HepG2 cells. ( I and J ) The effects of ANGPTL3 deficiency on the IL1R1–MyD88 and IL1R1–IL-1β interactions were determined by Co-IP ( I ) and quantitative analysis ( J ) in HepG2 cells. Quantification results were based on at least three experimental repeats. AN3, ANGPTL3; KO, knockout. ** P < 0.01; *** P < 0.001; ns, not significant.

Article Snippet: Antibodies against ANGPTL3 (R&D Systems, AF3829), MyD88 (Cell Signaling Technology, 4283), Flag tag (Sigma, F1804), HA tag (Sigma, H3663), and Myc tag (Cell Signaling Technology, 2276) were purchased.

Techniques: Over Expression, Co-Immunoprecipitation Assay, Knock-Out

Journal: Journal of Molecular Cell Biology

Article Title: ANGPTL3 negatively regulates IL-1 β -induced NF- κ B activation by inhibiting the IL1R1-associated signaling complex assembly

doi: 10.1093/jmcb/mjad053

Figure Lengend Snippet: Intracellular ANGPTL3 inhibits the IL1R1–IL1RAP interaction to attenuate IL-1β-induced signaling. ( A and B ) Full-length ANGPTL3 and its truncation mutants were overexpressed in HEK293T cells. The effects of the full-length ANGPTL3 and its truncation mutants on the IL1R1–IL1RAP interaction were determined by Co-IP ( A ) and quantitative analysis ( B ). ( C – F ) Wild-type ANGPTL3 and its loss-of-function mutants were overexpressed in HEK293T cells. ( C ) The effects of wild-type and mutant ANGPTL3 on the regulation of IL-1β-induced NF-κB activation were determined by the reporter assay (left panel). The expression levels of Flag-tagged wild-type and mutant ANGPTL3 in cell lysate and supernatant were detected by western blotting (right panel). ( D ) The interaction of overexpressed IL1R1 with wild-type and mutant ANGPTL3 was determined by Co-IP. ( E and F ) The effects of wild-type and mutant ANGPTL3 on the IL1R1–IL1RAP interaction was determined by Co-IP ( E ) and quantitative analysis ( F ). Quantification results were based on at least three experimental repeats. AN3, ANGPTL3; FL, full-length; Sup., supernatant; WT, wild-type. * P < 0.05; ** P < 0.01; *** P < 0.001; ns, not significant.

Article Snippet: Antibodies against ANGPTL3 (R&D Systems, AF3829), MyD88 (Cell Signaling Technology, 4283), Flag tag (Sigma, F1804), HA tag (Sigma, H3663), and Myc tag (Cell Signaling Technology, 2276) were purchased.

Techniques: Co-Immunoprecipitation Assay, Mutagenesis, Activation Assay, Reporter Assay, Expressing, Western Blot

Journal: Journal of Molecular Cell Biology

Article Title: ANGPTL3 negatively regulates IL-1 β -induced NF- κ B activation by inhibiting the IL1R1-associated signaling complex assembly

doi: 10.1093/jmcb/mjad053

Figure Lengend Snippet: Primers used in this study.

Article Snippet: Antibodies against ANGPTL3 (R&D Systems, AF3829), MyD88 (Cell Signaling Technology, 4283), Flag tag (Sigma, F1804), HA tag (Sigma, H3663), and Myc tag (Cell Signaling Technology, 2276) were purchased.

Techniques:

Journal: Journal of lipid research

Article Title: Decoding the role of angiopoietin-like protein 4/8 complex-mediated plasmin generation in the regulation of LPL activity.

doi: 10.1016/j.jlr.2023.100441

Figure Lengend Snippet: Fig. 1. ANGPTL4/8-mediated plasmin generation blocks ANGPTL3/8-mediated LPL inhibition. A: LPL stable expres- sion cells were incubated with lipase substrate after being pre- viously incubated with vehicle alone, ANGPTL4/8, tPA + plasminogen, or ANGPTL4/8 + tPA + plasminogen. LPL ac- tivity was calculated as a percent of vehicle control. Results are shown as the mean ± SD (n = 10 from 4 independent experi- ments) (*P < 0.001 vs. vehicle). B: LPL stable expression cells were incubated with vehicle alone throughout the experiment or in the absence or presence of ANGPTL3/8 after being previously incubated with vehicle alone, ANGPTL4/8 alone, tPA + plasminogen, or ANGPTL4/8 + tPA + plasminogen. Afterward, LPL activity was assessed following the addition of fluorescent lipase substrate and calculated as a percent of vehicle control. Results are shown as the mean ± SD (n = 8 from three independent experiments) (*P < 0.001 vs. ANGPTL3/8 alone). C: Samples from Figure 1B that contained ANGPTL3/8 were assessed using ANGPTL3 antibody Western blotting. Re- sults are representative of three independent experiments, with two replicates for each experiment (total n = 6 from three in- dependent experiments). ANGPTL, angiopoietin-like protein; LPL, lipoprotein lipase; tPA, tissue plasminogen activator.

Article Snippet: Human plasminogen (1939-SE), anti-ApoC2 rabbit antibody (MAF4497), anti-ANGPTL3 rabbit polyclonal antibody (AF3829), and anti-LPL antibody (AF7197) were purchased from R&D systems.

Techniques: Inhibition, Incubation, Control, Expressing, Activity Assay, Western Blot

Journal: Journal of lipid research

Article Title: Decoding the role of angiopoietin-like protein 4/8 complex-mediated plasmin generation in the regulation of LPL activity.

doi: 10.1016/j.jlr.2023.100441

Figure Lengend Snippet: Fig. 2. ANGPTL4/8-mediated plasmin generation blocks LPL inhibition by ANGPTL4 and ANGPTL3. A: LPL stable expression cells were incubated with vehicle alone throughout the experiment or in the absence or presence of ANGPTL4 after being previously preincubated with vehicle alone, ANGPTL4/8 alone, tPA + plasminogen, or ANGPTL4/8 + tPA + plasminogen. Afterward, LPL activity was assessed following the addition of fluorescent lipase substrate and calculated as a percent of vehicle control. Results are shown as the mean ± SD (n = 6 from two independent experiments) (*P < 0.001 vs. ANGPTL4 alone). B: LPL stable expression cells were incubated with vehicle alone throughout the experiment or in the absence or presence of ANGPTL3 after being previously preincubated with vehicle alone, ANGPTL4/8 alone, tPA + plasminogen, or ANGPTL4/8 + tPA + plasminogen. After- ward, LPL activity was assessed following the addition of fluo- rescent lipase substrate and calculated as a percent of vehicle control. Results are shown as the mean ± SD (n = 4 from two independent experiments) (*P < 0.001 vs. ANGPTL3 alone). C: Samples from Figure 2B that contained ANGPTL3 were

Article Snippet: Human plasminogen (1939-SE), anti-ApoC2 rabbit antibody (MAF4497), anti-ANGPTL3 rabbit polyclonal antibody (AF3829), and anti-LPL antibody (AF7197) were purchased from R&D systems.

Techniques: Inhibition, Expressing, Incubation, Activity Assay, Control

Journal: Journal of lipid research

Article Title: Decoding the role of angiopoietin-like protein 4/8 complex-mediated plasmin generation in the regulation of LPL activity.

doi: 10.1016/j.jlr.2023.100441

Figure Lengend Snippet: Fig. 5. Characterization of ANGPTL4/8-mediated plasmin generation and its effects on LPL. A: The ability of tPA to convert plasminogen to plasmin in the presence of ANGPTL4/8 was examined after addition of ANGPTL3/8, ANGPTL3, ANGPTL4, ApoC3, or ApoC2, with the molar ratio of each protein to ANGPTL4/8 being the same as in the LPL activity assays. Generation of plasmin was measured us- ing colorimetric plasmin activity assays, with absorbances recorded spectrophotometrically. Results shown are repre- sentative of three independent experiments. B: LPL stable–expressing cells were preincubated with ANGPTL4/8, followed by incubation in the absence or presence of tPA + plasminogen. Cells were washed, and heparin was added to release membrane-bound LPL, which was measured via Western blotting. Results are presentative of 6 replicates from 2 independent experiments. C: Following removal of membrane-bound LPL, cells were solubilized using RIPA buffer, and LPL present within the cells was analyzed via Western blotting. Results are presentative of six replicates from two independent experiments. ANGPTL, angiopoietin- like protein; LPL, lipoprotein lipase; tPA, tissue plasminogen activator.

Article Snippet: Human plasminogen (1939-SE), anti-ApoC2 rabbit antibody (MAF4497), anti-ANGPTL3 rabbit polyclonal antibody (AF3829), and anti-LPL antibody (AF7197) were purchased from R&D systems.

Techniques: Activity Assay, Expressing, Incubation, Membrane, Western Blot

Journal: Journal of lipid research

Article Title: Decoding the role of angiopoietin-like protein 4/8 complex-mediated plasmin generation in the regulation of LPL activity.

doi: 10.1016/j.jlr.2023.100441

Figure Lengend Snippet: Fig. 6. An updated model for adipose tissue LPL regulation in the postprandial state. After feeding, increased ANGPTL4/8 (a) binds LPL, protects it from inactivation by ANGPTL4, but also partially inhibits LPL (light blue color) (b). Partially active ANGPTL4/8-LPL binds GPIHBP1 on abluminal endothelial surfaces (c) and is translocated (d). Concurrently, tPA secreted by the endothelium and plasminogen present on endothelial plasminogen receptors (PLG-R) bind the LPL-ANGPTL4/8 complex on luminal capillary surfaces (e). ANGPTL4/8 increases tPA-mediated generation of plasmin, which cleaves ANGPTL4/8 (f) to restore LPL activity (dark blue color). The plasmin generated (g) also protects LPL from inhibition by circulating ANGPTL3/8, ANGPTL3, and ApoC3 as well as from any localized ANGPTL4 that may be present (for the purpose of clarity only ANGPTL3/8 is shown in the figure) (h), while permitting ApoC2-mediated LPL stimulation to occur (i). ANGPTL, angiopoietin-like protein; LPL, lipoprotein lipase; tPA, tissue plasminogen activator.

Article Snippet: Human plasminogen (1939-SE), anti-ApoC2 rabbit antibody (MAF4497), anti-ANGPTL3 rabbit polyclonal antibody (AF3829), and anti-LPL antibody (AF7197) were purchased from R&D systems.

Techniques: Activity Assay, Generated, Inhibition

Journal: Cell reports

Article Title: ANGPTL3 orchestrates hepatic fructose sensing and metabolism

doi: 10.1016/j.celrep.2025.115962

Figure Lengend Snippet: (A) Diagram of the experimental design. Male mice were fed with chow or MASLD diet for 6 weeks. Primary hepatocytes were isolated and cultured for 48 h. Conditioned media were collected, and AML12 cells were treated with conditioned media for 4 h, after which fructose uptake in AML12 cells was measured by [3H]-fructose incorporation. (B) Fructose uptake in AML12 cells treated with fractions of the conditioned medium separated by a 3K filter using >3 kDa (proteins) or <3 kDa (metabolites) ( N = 3 samples per group, repeated in two independent experiments). (C) Fractions of MASLD media separated by fast protein liquid chromatography (FPLC) fractionation and activity in inducing fructose uptake in AML12 cells ( N = 2 samples per fraction for the fructose uptake assays). (D) Silver stain of all fractions of MASLD media separated by FPLC fractionation. (E) Number of total proteins and secreted proteins identified by unbiased proteomics. (F) Venn diagram showing the number of proteins that overlap between datasets. (G) Fold change of differentially expressed genes in male chow and MASLD mouse livers measured by single-cell RNA sequencing. (H) Angptl3 levels in hepatocyte-conditioned media from male mice measured by ELISA ( N = 3 samples, repeated in two independent experiments). (I) Fructose uptake measured by [3H]-fructose incorporation in AML12 cells after 24-h treatments with 100 nM recombinant mouse Angptl3 ( N = 3 samples per group, repeated in three independent experiments), Angptl4, or Tsk ( N = 3 samples per group). (J) Fructose uptake measured by [3H]-fructose incorporation in AML12 cells after 24-h treatments with 100 nM recombinant mouse Angptl3 or human insulin ( N = 3 samples per group, repeated in two independent experiments). (K) Circulating male mouse Angptl3 levels measured by ELISA ( N = 5 mice per group). (L) Weekly body weights of male mice fed fructose or glucose in drinking water ( N = 5 mice per group). (M) Water consumption in male mice fed fructose or glucose in drinking water ( N = 5 mice per group). (N) Weekly blood glucose of male mice fed fructose or glucose in drinking water ( N = 5 mice per group). Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001 by two-tailed Student’s t test (I), one-way ANOVA (B, I, and J), or two-way ANOVA (K, L, M, and N).

Article Snippet: Angptl3 expression plasmid , OriGene , MR207262.

Techniques: Isolation, Cell Culture, Fast Protein Liquid Chromatography, Fractionation, Activity Assay, Silver Staining, RNA Sequencing, Enzyme-linked Immunosorbent Assay, Recombinant, Two Tailed Test

Journal: Cell reports

Article Title: ANGPTL3 orchestrates hepatic fructose sensing and metabolism

doi: 10.1016/j.celrep.2025.115962

Figure Lengend Snippet: (A) Relative gene expression analysis of angptl3 in male mouse organs ( N = 5 mice per group). (B) Relative gene expression analysis of khk (khk-a and khk-c) in male mouse organs ( N = 5 mice per group). (C) Relative gene expression analysis of aldob in male mouse organs ( N = 5 mice per group). (D–F) Gene expression levels of angptl3, aldob, and khk (khk-a and khk-c) in male chow and MASLD mouse livers measured by single-cell RNA sequencing ( N = 5 mice per group). (G) Relative gene expression analysis of angptl3 in livers from AAV8-Ctl- and AAV8-shAngptl3-treated male mice ( N = 5 mice per group, repeated in two independent cohorts). (H) Representative western blot of the liver from AAV8-Ctl- and AAV8-shAngptl3-treated male mice using an anti-Angptl3 antibody ( N = 4 mice per group). (I) Relative gene expression analysis of hepatic lipogenesis genes from AAV8-Ctl- and AAV8-shAngptl3-treated male mice ( N = 5 mice per group). (J) Uptake of [3H]-fructose-derived carbons in the liver of male mice fed with MASLD diet for 1 week ( N = 5 mice per group). (K) Relative gene expression levels of Glut8 in livers from AAV8-Ctl- and AAV8-shAngptl3-treated male mice ( N = 5 mice per group). (L) Relative gene expression levels of fructolysis enzyme genes in livers from AAV8-Ctl- and AAV8-shAngptl3-treated male mice ( N = 5 mice per group). (M) Representative western blot of fructolysis enzymes in livers from AAV8-Ctl- and AAV8-shAngptl3-treated male mice ( N = 5 mice per group). (N) Quantification of aldob protein levels relative to β-actin in (J) ( N = 5 mice per group). (O) Quantification of Khk protein levels relative to β-actin in (J) ( N = 5 mice per group). (P) Total cholesterol levels in the serum from AAV8-Ctl- and AAV8-shAngptl3-treated male mice ( N = 3–5 mice per group). (Q) Total triglycerol levels in the serum from AAV8-Ctl- and AAV8-shAngptl3-treated male mice ( N = 5 mice per group). (R) Water consumption in AAV8-Ctl- and AAV8-shAngptl3-treated male mice fed fructose in drinking water ( N = 3 mice per group). (S) In vivo lactic acid uptake in the liver measured by [14C]-lactate incorporation from male mice fed with MASLD diet for 1 week ( N = 11 mice per group). Data are presented as mean ± SEM. *p < 0.05, **p < 0.01, ***p < 0.001 by two-tailed Student’s t test (G, J, K, N, O, P, Q, R, and S) or two-way ANOVA (I and L).

Article Snippet: Angptl3 expression plasmid , OriGene , MR207262.

Techniques: Gene Expression, RNA Sequencing, Western Blot, Derivative Assay, In Vivo, Two Tailed Test

Journal: Cell reports

Article Title: ANGPTL3 orchestrates hepatic fructose sensing and metabolism

doi: 10.1016/j.celrep.2025.115962

Figure Lengend Snippet: (A) [3H]-fructose uptake in AML12 cells after treatment with increasing concentrations of LPL in the presence of 100 nM human ANGPTL3 ( N = 3 samples per group). (B) Schematic of generated point mutants of ANGPTL3. (C) [3H]-fructose uptake in AML12 cells after treatment with 100 nM wild-type or catalytically dead mutant of human ANGPTL3 in the presence or absence of LPL ( N = 3 samples per group). (D) Representative western blot and quantification of p -AKT, p -AKT substrate, p-S6 in HepG2 cells treated with indicated doses of full-length human ANGPTL3, and positive controls showing the intracellular signaling pathways over time ( N = 1 sample per group, repeated in two independent experiments). (E) Representative western blot and quantification of p -AKT, p -AKT substrate, p-S6 in HepG2 cells treated with indicated doses of C-terminal human ANGPTL3, and positive controls showing the intracellular signaling pathways over time ( N = 1 sample per group, repeated in two independent experiments). (F) Representative western blot of p -AKT; p -AKT substrate; p-S6 induced by C-terminal human ANGPTL3 (100 nM); or positive controls in HepG2 cells pretreated with PI3K inhibitors wortmannin (1 μM), mTORC1 inhibitor rapamycin (100 nM), mTORC1/2 dual inhibitor torin (100 nM), PKA inhibitor H89 (10 μM), or the translation inhibitor cycloheximide (10 μM) ( N = 1 sample per group). (G) [3H]-fructose uptake in AML12 cells after treatment with wortmannin with or without ANGPTL3 ( N = 8–9 samples per group). (H) Schematic of ANGPTL3-induced cellular signaling pathways in hepatocytes. Data are presented as mean ± SEM. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001 by one-way ANOVA (A, C, and G).

Article Snippet: Angptl3 expression plasmid , OriGene , MR207262.

Techniques: Generated, Mutagenesis, Western Blot, Protein-Protein interactions

Journal: Translational Oncology

Article Title: ANGPTL3 overcomes sorafenib resistance via suppression of SNAI1 and CPT1A in liver cancer

doi: 10.1016/j.tranon.2024.102250

Figure Lengend Snippet: ANGPTL3 is lower expressed in HCC tissues and correlated with good prognosis in HCC patients. (A) Venn diagram of differentially expressed genes identified in two online available datasets (GSE14520 and GSE94550). Levels of ANGPTL3, MTTP, AMBP, RELN and SLC4A3 in GSE14520 (B) and GSE94550 (C) were analyzed. Kaplan-Meier analysis of overall (D) and recurrence-free survival (E) based on the indicated gene expression in GSE14520. Median expression levels of those genes were used as the cutoff. Survival was analyzed using the log-rank test. (F) Expression levels and survival outcomes of ANGPTL3 in TCGA database were analyzed using the GEPIA tool ( http://gepia.cancer-pku.cn/ ). **, P < 0.01.

Article Snippet: Antibodies specific for ANGPTL3 (OriGene Technologies, Inc., Rockville, MD, USA, catalog no TA807389), SNAI1 (Cell Signaling Technology, Boston, MA, USA, catalog no #2532), CPT1A (Abclonal, Woburn, MA, USA, catalog no A5307) and GAPDH (Merck Millipore, Billerica, MA, USA, catalog no G8795) were used.

Techniques: Gene Expression, Expressing

Journal: Translational Oncology

Article Title: ANGPTL3 overcomes sorafenib resistance via suppression of SNAI1 and CPT1A in liver cancer

doi: 10.1016/j.tranon.2024.102250

Figure Lengend Snippet: ANGPTL3 expressions are lower expressed in sorafenib-resistant liver cancer cell lines. (A) Cell viability was assayed in parental liver cancer cells- and sorafenib resistant cell lines (HepG2SR, Huh7SR and J7SR)-treated with/without sorafenib. In addition, the IC50 values of sorafenib for the indicated cell lines were shown. (B) ANGPTL3 expression was measured in liver cancer cell lines, as indicated, via qRT-PCR analysis. GAPDH was used as endogenous control.

Article Snippet: Antibodies specific for ANGPTL3 (OriGene Technologies, Inc., Rockville, MD, USA, catalog no TA807389), SNAI1 (Cell Signaling Technology, Boston, MA, USA, catalog no #2532), CPT1A (Abclonal, Woburn, MA, USA, catalog no A5307) and GAPDH (Merck Millipore, Billerica, MA, USA, catalog no G8795) were used.

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Translational Oncology

Article Title: ANGPTL3 overcomes sorafenib resistance via suppression of SNAI1 and CPT1A in liver cancer

doi: 10.1016/j.tranon.2024.102250

Figure Lengend Snippet: ANGPTL3 suppresses cell migration and enhanced sorafenib-induced cell death. (A) ANGPTL3 expression in human hepatocyte (HH) and liver cancer cell lines was detected by western blot analysis. GAPDH served as endogenous control. The intensity of ANGPTL3 was quantified and normalized to GAPDH, with relative fold-changes compared to those in HepG2 cells. (B) Stable overexpression of ANGPTL3 was established in Huh7, J7, and Mahlavu cell lines, while stable knockdown of ANGPTL3 was achieved in the Hep3B cell line. The expression levels of ANGPTL3 in vector control (vc) and ANGPTL3-stable cell lines were assessed by western blot analysis. The ANGPTL3 signal was quantified and normalized to GAPDH, with relative fold-changes compared to the vc group. (C) The cell migration in ANGPTL3 stable cell lines was evaluated by Transwell migration assay. The migrated cells were stained by crystal violet and quantified by Image J software. (D) Conditioned medium (CM) from ANGPTL3-overexpressing Mahlavu cell cultures was collected and used for treating liver cancer cells as indicated. The cell migration of liver cancer cells was detected by Transwell migration assay. (E) The cell viability was determined by AlamarBlue cell viability reagent in ANGPTL3-overexpressing Huh7 and Mahlavu cell lines treated with/without sorafenib (5 μM). (F) The cell motility in the indicated liver cancer cell lines was evaluated by Transwell migration assay. The migrated cells were stained by crystal violet and quantified by Image J software.

Article Snippet: Antibodies specific for ANGPTL3 (OriGene Technologies, Inc., Rockville, MD, USA, catalog no TA807389), SNAI1 (Cell Signaling Technology, Boston, MA, USA, catalog no #2532), CPT1A (Abclonal, Woburn, MA, USA, catalog no A5307) and GAPDH (Merck Millipore, Billerica, MA, USA, catalog no G8795) were used.

Techniques: Migration, Expressing, Western Blot, Control, Over Expression, Knockdown, Plasmid Preparation, Stable Transfection, Transwell Migration Assay, Staining, Software

Journal: Translational Oncology

Article Title: ANGPTL3 overcomes sorafenib resistance via suppression of SNAI1 and CPT1A in liver cancer

doi: 10.1016/j.tranon.2024.102250

Figure Lengend Snippet: ANGPTL3 induces lipid peroxidation of liver cancer cell lines (A) The cell motility in the indicated liver cancer cell lines was evaluated by Transwell migration assay. The migrated cells were stained by crystal violet and quantified by Image J software. (B) The cell viability was determined by AlamarBlue cell viability reagent in ANGPTL3-overexpressing HepG2SR cell lines treated with/without sorafenib. The lipid peroxidation in ANGPTL3 stable cell lines (C) and sorafenib resistant cells (D) was assessed by lipid peroxidation assay. *, P < 0.05; **, P < 0.01.

Article Snippet: Antibodies specific for ANGPTL3 (OriGene Technologies, Inc., Rockville, MD, USA, catalog no TA807389), SNAI1 (Cell Signaling Technology, Boston, MA, USA, catalog no #2532), CPT1A (Abclonal, Woburn, MA, USA, catalog no A5307) and GAPDH (Merck Millipore, Billerica, MA, USA, catalog no G8795) were used.

Techniques: Transwell Migration Assay, Staining, Software, Stable Transfection, Peroxidation Assay

Journal: Translational Oncology

Article Title: ANGPTL3 overcomes sorafenib resistance via suppression of SNAI1 and CPT1A in liver cancer

doi: 10.1016/j.tranon.2024.102250

Figure Lengend Snippet: SNAI1 expression is negatively regulated by ANGPTL3 (A-B) MRNA levels of SNAI1 in ANGPTL3 stable cell lines were determined via qRT-PCR. GAPDH was used as endogenous control. (C) Protein levels of SNAI1 in the ANGPTL3-overexpressing Mahlavu cell lines were measured via western blot analysis. GAPDH was used as endogenous control. (D) The correlation between ANGPTL3 and SNAI1 in TCGA dataset was analyzed via Pearson correlation.

Article Snippet: Antibodies specific for ANGPTL3 (OriGene Technologies, Inc., Rockville, MD, USA, catalog no TA807389), SNAI1 (Cell Signaling Technology, Boston, MA, USA, catalog no #2532), CPT1A (Abclonal, Woburn, MA, USA, catalog no A5307) and GAPDH (Merck Millipore, Billerica, MA, USA, catalog no G8795) were used.

Techniques: Expressing, Stable Transfection, Quantitative RT-PCR, Control, Western Blot

Journal: Translational Oncology

Article Title: ANGPTL3 overcomes sorafenib resistance via suppression of SNAI1 and CPT1A in liver cancer

doi: 10.1016/j.tranon.2024.102250

Figure Lengend Snippet: ANGPTL3 facilitates CPT1A protein degradation in sorafenib-resistant liver cancer cell lines. (A) The protein levels of CPT1A in the indicated cell lines were measured via western blot analysis. vc: vector control group. GAPDH was used as endogenous control. The fold-changes of CPT1A in the specified cell lines were quantified and presented. (B) CPT1A expression in ANGPTL3-overexpressing cell lines, as indicated, with or without MG132 treatment was detected via western blot analysis. GAPDH was used as endogenous control. The fold-changes of CPT1A in the specified cell lines were quantified and presented. (C) The turnover rate of CPT1A protein was detected in cycloheximide (CHX) chase experiments. The intensity of CPT1A was calculated and normalized to the endogenous control, GAPDH.

Article Snippet: Antibodies specific for ANGPTL3 (OriGene Technologies, Inc., Rockville, MD, USA, catalog no TA807389), SNAI1 (Cell Signaling Technology, Boston, MA, USA, catalog no #2532), CPT1A (Abclonal, Woburn, MA, USA, catalog no A5307) and GAPDH (Merck Millipore, Billerica, MA, USA, catalog no G8795) were used.

Techniques: Western Blot, Plasmid Preparation, Control, Expressing