Journal: Science signaling

Article Title: Designer high-density lipoprotein particles enhance endothelial barrier function and suppress inflammation

doi: 10.1126/scisignal.adg9256

Figure Lengend Snippet: (A) Side view of A1M-S1P complex with and without the phospholipids, highlighting the S1P binding pocket in both the ApoM-S1P and ApoA1-fused systems. Monomer 1 - ApoA1 in sienna, linker in brown, and ApoM in beige. Monomer 2 - ApoA1 in sage, linker in turquoise, and ApoM in teal. Detail of the binding pocket in A1M-S1P and ApoM-S1P are shown in the right panel. (B) Projection of protein atoms along the top two dominant eigenvectors (eigenvector 1 - PC1 and eigenvector 2 – PC2) is responsible for over 40% of the dominant collective variances in the structure set, generating 5 clusters for which the three-dimensional structure of the most representative member of each cluster is shown. Permanence times: cluster 1 – 9.6%, cluster 2 – 52.9%, cluster 3 – 18.7%, cluster 4 – 12.2%, cluster 5 – 6.5%. (C) Distribution of the proportion of variance between the PCs.

Article Snippet: The A1M fusion was constructed using plasmids for murine ApoA1 (MR203500) and murine ApoM (MR201811) obtained from OriGene.

Techniques: Binding Assay

Journal: Science signaling

Article Title: Designer high-density lipoprotein particles enhance endothelial barrier function and suppress inflammation

doi: 10.1126/scisignal.adg9256

Figure Lengend Snippet: (A) HMEC-1 cells expressing an NF-κB-luciferase reporter were assayed for TNFα-induced NF-κB reporter activity in the presence of ApoA1, A1M, A1M-S1P, and ApoM-Fc-S1P (N = 3 biological replicates per group). Data are presented as means + S.D. **P < 0.01, ***P < 0.001, ****P < 0.0001 by ordinary two-way ANOVA with Tukey’s multiple comparisons test. TNFα is the reference group. (B) HUVECs were starved for 1 h, pre-treated for 10 minutes with ApoM-Fc-S1P (100 nM), iloprost (200 nM), both ApoM-Fc-S1P and iloprost, A1M (200 μg/mL), A1M-S1P (200 μg/mL), or A1M-iloprost (200 μg/mL) and induced with TNFα (10 ng/mL) for 5 hours. Lysates were subjected to immunoblot analysis for ICAM-1. Quantification of immunoblots was analyzed from 3 biological replicates per group. Data are presented as means ± S.D. *P < 0.05, **P < 0.01, ****P < 0.0001 by ANOVA with post-hoc Holm-Sidak’s multiple comparisons test. (C) Cholesterol efflux in response to human HDL (hHDL), human ApoA1 protein (hApoA1), and bacterial A1M (bA1M) in PMA-induced THP-1 cells (N ≥ 5 independent experiments). Data are presented as means + S.D. *P < 0.05, ***P < 0.001, ****P < 0.0001 by one-way ANOVA with Dunnett’s multiple comparisons test.

Article Snippet: The A1M fusion was constructed using plasmids for murine ApoA1 (MR203500) and murine ApoM (MR201811) obtained from OriGene.

Techniques: Expressing, Luciferase, Activity Assay, Western Blot

Journal: Cell Death Discovery

Article Title: APOA2-mediated endothelial mesenchymal transition and cancer lipid metabolism reprogramming confers antiangiogenic drug resistance through TGF-β

doi: 10.1038/s41420-026-02984-5

Figure Lengend Snippet: a MRI images, changes in AFP, and quantification of tumor volume in representative HCC-resistant patients before and after apatinib treatment. Scale bars, 10 cm. b MRI images, changes in AFP, and quantification of tumor thrombus volume in representative HCC-sensitive patients before and after apatinib treatment. Scale bars, 10 cm. c The GO pathway analysis of differential genes was enriched, with APOA2 ranking first in the small molecule metabolic process. d The KEGG pathway analysis of differential genes was enriched, with APOA2 ranking first in the PPAR signaling pathway. e Intensity of APOA2 IHC staining in 10 paired HCC patient tissues (sensitive vs resistant). Scale bars, 50 µm. f Predicted network diagram of the interactions between APOA2 and VEGFR2.

Article Snippet: Primary APOA2 antibody (Proteintech, 16845-1-AP, CHN) was diluted at 1:50, while mouse/rabbit linker (Santa Cruz, sc2357, USA) was diluted at 1:100.

Techniques: Immunohistochemistry

Journal: Cell Death Discovery

Article Title: APOA2-mediated endothelial mesenchymal transition and cancer lipid metabolism reprogramming confers antiangiogenic drug resistance through TGF-β

doi: 10.1038/s41420-026-02984-5

Figure Lengend Snippet: After the evaluation of acquired drug resistance, the HCC-bearing mice were divided into 3 groups: (C)control ( n = 7), (R)resistant ( n = 7), and (S)sensitive ( n = 24). a Quantification of subcutaneous tumor mass and volume in the three groups. Numbers 1–7 represent control group, 1–7 for resistance group, and 1–7 for sensitivity group. * p < 0.05, ns, not significant. b . Representative images of CD31 immunofluorescence staining of tumor tissue. The upper part of the image is a comprehensive view of CD31 staining of the entire tumor tissue section, while the bottom part shows CD31 staining under magnification. CD31 + microvessel signals were randomly quantified from 12 fields ( n = 7 mice in each group), Scale bars, 50 µm. * p < 0.05, ns, not significant. c and d . Western blotting of APOA2, VEGFR2, and P-VEGFR2 in tumor tissue ( n = 4). e Representative images of APOA2 IHC staining intensity of tumor tissue, Scale bars, 100 µm. The upper part of the image is a comprehensive view of APOA2 staining of the entire tumor tissue section, while the bottom part shows APOA2 staining under magnification. Scale bars, 100 µm. * p < 0.05, ns, not significant.

Article Snippet: Primary APOA2 antibody (Proteintech, 16845-1-AP, CHN) was diluted at 1:50, while mouse/rabbit linker (Santa Cruz, sc2357, USA) was diluted at 1:100.

Techniques: Control, Immunofluorescence, Staining, Western Blot, Immunohistochemistry

Journal: Cell Death Discovery

Article Title: APOA2-mediated endothelial mesenchymal transition and cancer lipid metabolism reprogramming confers antiangiogenic drug resistance through TGF-β

doi: 10.1038/s41420-026-02984-5

Figure Lengend Snippet: a Representative images of HCC cell lines overexpressing APOA2. Scale bars, 100 µm. Western blotting verified the protein levels of APOA2, VEGFR2, and P-VEGFR2 in 4 cell lines. b – d Subcutaneous tumor, tumor volume and tumor mass quantification, along with tumor growth curve of each group ( n = 6). e – g Characterization of tumor inhibition rate in each group. *** p < 0.001, **** p < 0.0001, ns not significant.

Article Snippet: Primary APOA2 antibody (Proteintech, 16845-1-AP, CHN) was diluted at 1:50, while mouse/rabbit linker (Santa Cruz, sc2357, USA) was diluted at 1:100.

Techniques: Western Blot, Inhibition

Journal: Cell Death Discovery

Article Title: APOA2-mediated endothelial mesenchymal transition and cancer lipid metabolism reprogramming confers antiangiogenic drug resistance through TGF-β

doi: 10.1038/s41420-026-02984-5

Figure Lengend Snippet: a – f The experiment comprised two groups: control (Control) and APOA2 overexpression (APOA2). a , d Colony formation assay conducted on MHCC97H and SMMC7721 cells ( n = 3), b , e Representative images from EdU assay performed on MHCC97H and SMMC7721 cells, with EdU signals quantified from 12 fields ( n = 6), Scale bars, 50 µm. c , f CCK8 assay results of MHCC97H and SMMC7721 cells. g The experiment comprised 4 groups: control group receiving solvent and apatinib (Control + Vehicle vs Control + Apatinib), and the APOA2 overexpression group receiving solvent and apatinib (APOA2+Vehicle vs APOA2+Apatinib). Representative images of Ki67 and cleaved-caspase 3 immunofluorescence staining of tumor tissue, with Ki67+ and cleaved-caspase 3+ signals quantified from 12 fields ( n = 6). Scale bars, 100 µm. * p < 0.05, ns not significant.

Article Snippet: Primary APOA2 antibody (Proteintech, 16845-1-AP, CHN) was diluted at 1:50, while mouse/rabbit linker (Santa Cruz, sc2357, USA) was diluted at 1:100.

Techniques: Control, Over Expression, Colony Assay, EdU Assay, CCK-8 Assay, Solvent, Immunofluorescence, Staining

Journal: Cell Death Discovery

Article Title: APOA2-mediated endothelial mesenchymal transition and cancer lipid metabolism reprogramming confers antiangiogenic drug resistance through TGF-β

doi: 10.1038/s41420-026-02984-5

Figure Lengend Snippet: a Representative bright-field images of tubular formation in HUVECs cultured with conditioned media from MHCC97H and SMCC7721 (overexpressing APOA2 or not). Scale bar, 400 µm. b Representative images of EdU assay conducted on HUVECs cultured with conditioned media from MHCC97H and SMCC7721 (overexpressing APOA2 or not), with EdU signals quantified from 12 fields ( n = 6). Scale bars, 100 µm. c Representative images of migration of HUVECs cultured with conditioned media from MHCC97H and SMCC7721 (overexpressing APOA2 or not), Scale bars, 100 µm. * p < 0.05, ns not significant.

Article Snippet: Primary APOA2 antibody (Proteintech, 16845-1-AP, CHN) was diluted at 1:50, while mouse/rabbit linker (Santa Cruz, sc2357, USA) was diluted at 1:100.

Techniques: Cell Culture, EdU Assay, Migration

Journal: Cell Death Discovery

Article Title: APOA2-mediated endothelial mesenchymal transition and cancer lipid metabolism reprogramming confers antiangiogenic drug resistance through TGF-β

doi: 10.1038/s41420-026-02984-5

Figure Lengend Snippet: a Volcano plot of differentially expressed proteins between the APOA2 overexpression group and the control group, with a fold change ≥ 1.5 and adjusted p < 0.01 ( n = 3). Gray denotes no significant difference, blue indicates low expression, and red indicates high expression. The volcano plot labels the five highest expressed proteins and five lowest expressed proteins. b Histogram of 5 highest expressed proteins and 5 lowest expressed proteins ( n = 3). c Enrichment analysis of KEGG pathways for differentially expressed proteins between the APOA2 overexpression group and the control group. d Histogram showing levels of TGF-β ligand, receptor, and signaling pathway-related protein in proteomics (n = 3). e Pearson correlation analysis of five highly expressed proteins and TGF-β in HCC. f Relative mRNA levels of TGF-β in HCC cells and tumor tissues with or without overexpressing APOA2 analyzed by real-time quantitative PCR ( n = 6). g ELISA detection of TGF-β in HCC cell (with or without overexpressing APOA2) medium and serum of HCC-bearing mice (with or without overexpressing APOA2). h Western blotting detecting the protein levels of TGF-βR1, TGF-βR2, P-TGFβR1 and P-TGFβR2 in HCC cells and tumor tissues with or without overexpressing APOA2. * p < 0.05, ns, not significant.

Article Snippet: Primary APOA2 antibody (Proteintech, 16845-1-AP, CHN) was diluted at 1:50, while mouse/rabbit linker (Santa Cruz, sc2357, USA) was diluted at 1:100.

Techniques: Over Expression, Control, Expressing, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Cell Death Discovery

Article Title: APOA2-mediated endothelial mesenchymal transition and cancer lipid metabolism reprogramming confers antiangiogenic drug resistance through TGF-β

doi: 10.1038/s41420-026-02984-5

Figure Lengend Snippet: a Enrichment analysis of GO pathways for differentially expressed proteins between APOA2 overexpression and the control group, with a fold change ≥ 1.5 and adjusted p -value < 0.01. b Glucose consumption in MHCC97H cells in the designated group ( n = 6). c Lactate production in MHCC97H cells in the designated group ( n = 6). d Mito Fuel Flex test revealed dependence on the oxidation of glucose, glutamine, or FA in MHCC97H cells in the designated group ( n = 6). e BODIPY-FA uptake in MHCC97H cells in the designated group. f β-oxidation assay in MHCC97H cells in the designated group ( n = 6). g Intracellular ATP levels in MHCC97H cells in the designated group ( n = 6). * p < 0.05, ns not significant.

Article Snippet: Primary APOA2 antibody (Proteintech, 16845-1-AP, CHN) was diluted at 1:50, while mouse/rabbit linker (Santa Cruz, sc2357, USA) was diluted at 1:100.

Techniques: Over Expression, Control, Oxidation Assay

Journal: Journal of Translational Medicine

Article Title: Characterization of the proteome of stable and unstable carotid atherosclerotic plaques using data-independent acquisition mass spectrometry

doi: 10.1186/s12967-023-04723-1

Figure Lengend Snippet: Validation of ferroptosis- and lipid metabolism-associated DEPs using IHC in a validation cohort. A Representative images (see Additional file : Figure S4a,b for 500 µm images) of immunohistochemical staining of TFR1, TF, AIFM2, DPP4, and GCLC proteins in stable and unstable plaques in the plaque fibrous cap region (black arrows) and immunohistochemical staining for SLC1A5, BID, and APOA5 proteins in the plaque lipid core region (black arrows). B In unstable plaques, the levels of TFR1, TF, AIFM2, DPP4, GCLC, SLC1A5, BID, and APOA5 were significantly increased, while other differences were not statistically significant. IHC immunohistochemistry. TFR1 transferrin receptor protein 1; TF transferrin; AIFM2 apoptosis-inducing factor 2; DPP4 dipeptidyl peptidase 4; GCLC glutamate-cysteine ligase catalytic. SLC1A5 solute carrier family 1, member 5; BID BH3 interacting-domain death agonist; APOA5, apolipoprotein A-V; CETP cholesteryl ester transfer protein. *P < 0.05; **P < 0.01 Student’s t test (two-tailed distribution)

Article Snippet: Primary antibodies for IHC against solute carrier family 1, member 5 (SLC1A5) (rabbit polyclonal, 20350-1-AP), apoptosis-inducing factor 2 (AIFM2) (rabbit polyclonal, 20886–1-AP), BH3 interacting-domain death agonist (BID) (rabbit polyclonal, 10988-1-AP), dipeptidyl peptidase 4 (DPP4) (rabbit polyclonal, 29403-1-AP), transferrin receptor protein 1 (TFR1) (mouse monoclonal, 66180–1-Ig), transferrin (TF) (rabbit polyclonal, 17435-1-AP), glutamate–cysteine ligase catalytic (GCLC) (rabbit polyclonal, 12601–1-AP), cholesteryl ester transfer protein (CETP) (rabbit polyclonal, 13459-1-AP), and apolipoprotein A-V (APOA5) (rabbit polyclonal, 18019-1-AP) were purchased from Proteintech (China).

Techniques: Biomarker Discovery, Immunohistochemical staining, Staining, Immunohistochemistry, Two Tailed Test

Journal: Journal of Translational Medicine

Article Title: Characterization of the proteome of stable and unstable carotid atherosclerotic plaques using data-independent acquisition mass spectrometry

doi: 10.1186/s12967-023-04723-1

Figure Lengend Snippet: The differently expressed proteins (DEPs) that were validated using immunohistochemistry (all up regulated) (unstable/stable)

Article Snippet: Primary antibodies for IHC against solute carrier family 1, member 5 (SLC1A5) (rabbit polyclonal, 20350-1-AP), apoptosis-inducing factor 2 (AIFM2) (rabbit polyclonal, 20886–1-AP), BH3 interacting-domain death agonist (BID) (rabbit polyclonal, 10988-1-AP), dipeptidyl peptidase 4 (DPP4) (rabbit polyclonal, 29403-1-AP), transferrin receptor protein 1 (TFR1) (mouse monoclonal, 66180–1-Ig), transferrin (TF) (rabbit polyclonal, 17435-1-AP), glutamate–cysteine ligase catalytic (GCLC) (rabbit polyclonal, 12601–1-AP), cholesteryl ester transfer protein (CETP) (rabbit polyclonal, 13459-1-AP), and apolipoprotein A-V (APOA5) (rabbit polyclonal, 18019-1-AP) were purchased from Proteintech (China).

Techniques: Immunohistochemistry, Biomarker Discovery, Significance Assay

Journal: Journal of Translational Medicine

Article Title: Characterization of the proteome of stable and unstable carotid atherosclerotic plaques using data-independent acquisition mass spectrometry

doi: 10.1186/s12967-023-04723-1

Figure Lengend Snippet: Hypothetical characterization of altered molecular mechanisms in cells in the fibrous cap and lipid core regions of unstable carotid plaques. The expression of key proteins of ferroptosis and lipid metabolism is significantly increased in patients with unstable plaques, and there are mechanisms associated with both the promotion and inhibition of iron death. This possibly indicates that cells in different regions of the plaque are regulated by key proteins of ferroptosis that subsequently lead to increased plaque instability and expansion of the necrotic core. TFR1 transferrin receptor protein 1; TF transferrin; AIFM2 apoptosis-inducing factor 2; DPP4 dipeptidyl peptidase 4; GCLC glutamate-cysteine ligase catalytic; SLC1A5 solute carrier family 1, member 5; BID BH3 interacting-domain death agonist; APOA5 apolipoprotein A-V; CETP cholesteryl ester transfer protein; GPX4 glutathione peroxidase 4; GSH glutathione; CoQ10 coenzyme Q10; ROS reactive oxygen species; HDL high-density lipoprotein; ox-LDL oxidized low-density lipoprotein

Article Snippet: Primary antibodies for IHC against solute carrier family 1, member 5 (SLC1A5) (rabbit polyclonal, 20350-1-AP), apoptosis-inducing factor 2 (AIFM2) (rabbit polyclonal, 20886–1-AP), BH3 interacting-domain death agonist (BID) (rabbit polyclonal, 10988-1-AP), dipeptidyl peptidase 4 (DPP4) (rabbit polyclonal, 29403-1-AP), transferrin receptor protein 1 (TFR1) (mouse monoclonal, 66180–1-Ig), transferrin (TF) (rabbit polyclonal, 17435-1-AP), glutamate–cysteine ligase catalytic (GCLC) (rabbit polyclonal, 12601–1-AP), cholesteryl ester transfer protein (CETP) (rabbit polyclonal, 13459-1-AP), and apolipoprotein A-V (APOA5) (rabbit polyclonal, 18019-1-AP) were purchased from Proteintech (China).

Techniques: Expressing, Inhibition