Journal: Cell Communication and Signaling : CCS

Article Title: Hormone-sensitive lipase drives pro-resolving macrophage polarization and enhances efferocytosis

doi: 10.1186/s12964-025-02631-z

Figure Lengend Snippet: Genetic knockdown of HSL replicates the effects of pharmacological inhibition, thereby confirming its essential role in sustaining the M2 phenotype. Human M0 macrophages were transfected with control non-targeting siRNA or siRNA targeting HSL. (A) Efferocytosis efficiency was measured 48 h post-transfection ( n = 6 independent donors). (B) Secreted levels of the pro-resolving mediator Annexin A1 and the pro-inflammatory cytokine TNFα in culture supernatants, measured by ELISA ( n = 9–10 independent donors). (C) Relative mRNA expression of M2-associated genes (PPARγ, CD163, CD36, CD200R1) analyzed by qRT-PCR. ( n = 5, independent donors). (D) Relative mRNA expression of M1-associated genes (TNFα, IL1β, HIF1α) and HSL/ LIPE ( n = 5, independent donors). (E) Efferocytosis was measured in murine M0 bone marrow-derived macrophages (BMDMs) following siRNA-mediated knockdown of HSL ( n = 6 biological replicates). For panels A, B, and E, a paired t-test was used. For panels C and D, a two-way ANOVA was used. Data in A, B, and E are presented as box-and-whisker plots. Panels C & D show paired individual donors. * P < 0.05, ** P < 0.01, *** P < 0.001

Article Snippet: For siRNA-mediated gene silencing, M0-human macrophages or BMDMs were transfected with 25 nM of Silencer pre-designed siRNA targeting human LIPE /HSL (143993 and 143994, Life Technologies), Lipe mouse pre-designed siRNA Set A (HY-RS18621, MCE), or a non-targeting control pool (Dharmacon, Lafayette, CO) using Lipofectamine LTX Transfection Reagent (Thermo Fisher Scientific).

Techniques: Knockdown, Inhibition, Transfection, Control, Enzyme-linked Immunosorbent Assay, Expressing, Quantitative RT-PCR, Derivative Assay, Whisker Assay

Journal: Frontiers in Immunology

Article Title: Identification of a ubiquitin-binding domain protein, CD2AP, in predicting the prognosis and treatment of lung adenocarcinoma

doi: 10.3389/fimmu.2025.1726531

Figure Lengend Snippet: Analysis of proteomics data in the CPTAC database. (A) Comparison of the protein levels between normal (green) and tumour (red) samples. (B) Kaplan–Meier curve for each protein. (C) Pan-cancer analysis for CD2AP (green: normal tissue, red: cancer tissue). (D) Relationship between CD2AP protein level and immune infiltration in pan-cancer.

Article Snippet: CD2AP Human Pre-designed siRNA was acquired from MedChemExpress (MCE).

Techniques: Comparison

Journal: Frontiers in Immunology

Article Title: Identification of a ubiquitin-binding domain protein, CD2AP, in predicting the prognosis and treatment of lung adenocarcinoma

doi: 10.3389/fimmu.2025.1726531

Figure Lengend Snippet: TMB and immune cell analysis. (A) The top 20 mutated genes in the CD2AP -low subgroup. (B) The top 20 mutated genes in the CD2AP -high subgroup. In both panels, the genes are ordered from top to bottom by their mutation frequency. The most frequently mutated genes in the entire cohort are TP53 and TTN. (C) The infiltration fraction for each immune cell between the CD2AP -low(blue) and the CD2AP -high(red) subgroups. (D) The relationship between CD2AP copy number variation and the infiltration level of immune cells. *, P <0.05**, P <0.01; ***, P <0.001.

Article Snippet: CD2AP Human Pre-designed siRNA was acquired from MedChemExpress (MCE).

Techniques: Cell Analysis, Mutagenesis

Journal: Frontiers in Immunology

Article Title: Identification of a ubiquitin-binding domain protein, CD2AP, in predicting the prognosis and treatment of lung adenocarcinoma

doi: 10.3389/fimmu.2025.1726531

Figure Lengend Snippet: Pathway enrichment analysis. (A) GO analysis for the DEGs between the CD2AP -low and CD2AP -high subgroups. (B) KEGG analysis for the DEGs between the CD2AP -low and CD2AP -high subgroups. (C) GSEA analysis for CD2AP -low and CD2AP -high subgroups.

Article Snippet: CD2AP Human Pre-designed siRNA was acquired from MedChemExpress (MCE).

Techniques:

Journal: Frontiers in Immunology

Article Title: Identification of a ubiquitin-binding domain protein, CD2AP, in predicting the prognosis and treatment of lung adenocarcinoma

doi: 10.3389/fimmu.2025.1726531

Figure Lengend Snippet: Single-cell and IHC analysis. (A) Single-cell analysis for GSE99254 . (B) Healthy lung tissue (Patient ID: 2101) staining with CD2AP. (C) LUAD tissue (Patient ID: 2438) staining with CD2AP.

Article Snippet: CD2AP Human Pre-designed siRNA was acquired from MedChemExpress (MCE).

Techniques: Single-cell Analysis, Staining

Journal: Frontiers in Immunology

Article Title: Identification of a ubiquitin-binding domain protein, CD2AP, in predicting the prognosis and treatment of lung adenocarcinoma

doi: 10.3389/fimmu.2025.1726531

Figure Lengend Snippet: Exploration of CD2AP functions. (A) CD2AP gene expression across all cell types; (B) CD2AP gene expression in monocyte subtypes; (C) Expression distribution of CD2AP in cancer cells and monocytes; (D) Cellular interaction network of CD2AP + cancer cells LUAD; (E) Dot plot for the enrichment of ligand-receptor pathways; (F) KEGG pathway enrichment analysis for CD2AP + and CD2AP - monocytes.

Article Snippet: CD2AP Human Pre-designed siRNA was acquired from MedChemExpress (MCE).

Techniques: Gene Expression, Expressing

Journal: Frontiers in Immunology

Article Title: Identification of a ubiquitin-binding domain protein, CD2AP, in predicting the prognosis and treatment of lung adenocarcinoma

doi: 10.3389/fimmu.2025.1726531

Figure Lengend Snippet: Drug sensitivity analysis and molecular docking. (A) Venn plot indicating the potential targeted compounds. (B) The structures of CD2AP. (C) Molecular docking between afatinib and CD2AP. (D) Molecular docking between dasatinib and CD2AP.

Article Snippet: CD2AP Human Pre-designed siRNA was acquired from MedChemExpress (MCE).

Techniques:

Journal: Frontiers in Immunology

Article Title: Identification of a ubiquitin-binding domain protein, CD2AP, in predicting the prognosis and treatment of lung adenocarcinoma

doi: 10.3389/fimmu.2025.1726531

Figure Lengend Snippet: Histological experiments to validate the expression of CD2AP. (A) Validation of CD2AP expression in cancer and adjacent tissues by immunofluorescence (n=3 biologically independent samples); (B) Determine CD2AP protein levels using Western blot analysis; (C) Statistical chart for WB, data are presented as mean ± SD (n=5 biologically independent samples). * P < 0.05; (D) Results of rt-qPCR for 5 controls vs. 5 LUADs, data are presented as mean ± SD (n=5 biologically independent samples). ** P < 0.01.

Article Snippet: CD2AP Human Pre-designed siRNA was acquired from MedChemExpress (MCE).

Techniques: Expressing, Biomarker Discovery, Immunofluorescence, Western Blot, Quantitative RT-PCR

Journal: Frontiers in Immunology

Article Title: Identification of a ubiquitin-binding domain protein, CD2AP, in predicting the prognosis and treatment of lung adenocarcinoma

doi: 10.3389/fimmu.2025.1726531

Figure Lengend Snippet: Cell experiments to validate the function of CD2AP. (A) WB to show the knockdown efficiency of CD2AP; (B) Statistical chart for WB, data are presented as mean ± SD (n=3 biologically independent samples). ** P < 0.01; (C) Proliferative capacity of A549 cells by CCK-8 assay, data are presented as mean ± SD (n=3 technical replicates per group, representative of three independent experiments); (D) Cell migration capabilities by transwell; (E) Statistical chart for transwell counts, data are presented as mean ± SD (n=3 biologically independent samples). **** P < 0.0001; (F) Cell migration capabilities by wound healing assays; (G) Statistical chart for wound healing assays, data are presented as mean ± SD (n=3 biologically independent samples). ** P < 0.01.

Article Snippet: CD2AP Human Pre-designed siRNA was acquired from MedChemExpress (MCE).

Techniques: Knockdown, CCK-8 Assay, Migration

Journal: bioRxiv

Article Title: Curcumin Analogues Trigger HMOX1-Mediated Ferroptosis to Halt Endometrial Cancer Growth

doi: 10.1101/2025.11.18.689080

Figure Lengend Snippet: (A) Real-time PCR analysis of HMOX1 mRNA expression in KLE cells treated with 2 µM AKT-100, 5 µM HO-3867, or 30 µM hemin for 24 hours. (B) Real-time PCR analysis of HMOX1 mRNA expression in Hec50co cells treated with 0.5 µM AKT-100, 3 µM HO-3867, or 20 µM hemin for 24 hours. (C) Immunoblotting and densitometric quantification of HMOX1 protein levels in KLE cells under the same treatment conditions. (D) Immunoblotting and densitometric quantification of HMOX1 protein levels in Hec50co cells under the same treatment conditions. Data are presented as mean ± SD. p < 0.05, p < 0.01, p < 0.001; one-way ANOVA followed by Dunnett’s multiple comparisons test.

Article Snippet: HMOX1 Human Pre-designed siRNA Set A (MedChemExpress, #HY-RS06249) was combined with DreamFect Gold Transfection Reagent (OzBiosciences, #DG81000) and incubated for 5 minutes at room temperature.

Techniques: Real-time Polymerase Chain Reaction, Expressing, Western Blot

Journal: bioRxiv

Article Title: Curcumin Analogues Trigger HMOX1-Mediated Ferroptosis to Halt Endometrial Cancer Growth

doi: 10.1101/2025.11.18.689080

Figure Lengend Snippet: (A) Graphical illustration showing how novel curcumin analogues induce ferroptosis via heme oxygenase-1 (HMOX1) upregulation to suppress endometrial cancer growth. (B) FerroOrange staining assessing intracellular iron levels in KLE cells treated with 2 µM AKT-100 for 24 hours. (C) DCFH-DA staining measuring total ROS levels under the same treatment conditions. Data are presented as mean ± SD. p < 0.05, p < 0.01; unpaired t-test.

Article Snippet: HMOX1 Human Pre-designed siRNA Set A (MedChemExpress, #HY-RS06249) was combined with DreamFect Gold Transfection Reagent (OzBiosciences, #DG81000) and incubated for 5 minutes at room temperature.

Techniques: Analogues, Staining

Journal: bioRxiv

Article Title: Curcumin Analogues Trigger HMOX1-Mediated Ferroptosis to Halt Endometrial Cancer Growth

doi: 10.1101/2025.11.18.689080

Figure Lengend Snippet: CyQUANT cell proliferation assays of KLE cells treated with 2 µM AKT-100 in combination with: (A) 5 µM zinc protoporphyrin IX (ZnPP, HMOX1 inhibitor), (B) 50 nM liproxstatin-1 (ferroptosis inhibitor), or (C) 10 µM Z-VAD-FMK (pan-caspase/apoptosis inhibitor) for 72 hours. Data are presented as mean ± SD. p < 0.05, * p < 0.01, *** p < 0.0001; two-way ANOVA followed by Šídák’s multiple comparisons test.

Article Snippet: HMOX1 Human Pre-designed siRNA Set A (MedChemExpress, #HY-RS06249) was combined with DreamFect Gold Transfection Reagent (OzBiosciences, #DG81000) and incubated for 5 minutes at room temperature.

Techniques: CyQUANT Assay

Journal: bioRxiv

Article Title: Curcumin Analogues Trigger HMOX1-Mediated Ferroptosis to Halt Endometrial Cancer Growth

doi: 10.1101/2025.11.18.689080

Figure Lengend Snippet: (A) Immunoblotting and densitometric quantification of HMOX1 protein levels in KLE cells transfected with 100 nM negative control siRNA (siRNA-Neg) or HMOX-1–specific siRNA (siRNA-HMOX1) for 48 hours, followed by treatment with 2 µM AKT-100 for 24 hours. Data were analyzed using two-way ANOVA followed by Šídák’s multiple comparisons test. (B) CyQUANT cell proliferation assay of KLE cells transfected with 100 nM siRNA for 48 hours and then treated with 2 µM AKT-100 for 72 hours. Data were analyzed using one-way ANOVA followed by Tukey’s multiple comparisons test. (C) Representative images of colony formation in KLE cells transfected with 100 nM siRNA for 48 hours and treated with 2 µM AKT-100 for 24 hours. (D) Quantification of colony formation shown in (C). Data were analyzed using one-way ANOVA followed by Tukey’s multiple comparisons test. Data are presented as mean ± SD. p < 0.05, * p < 0.01, ** p < 0.001; ns, not significant.

Article Snippet: HMOX1 Human Pre-designed siRNA Set A (MedChemExpress, #HY-RS06249) was combined with DreamFect Gold Transfection Reagent (OzBiosciences, #DG81000) and incubated for 5 minutes at room temperature.

Techniques: Western Blot, Transfection, Negative Control, CyQUANT Assay, Proliferation Assay