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: 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: The sections were then incubated overnight at 4 °C with a primary antibody against CD2AP (Proteintech, Wuhan, China).

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: The sections were then incubated overnight at 4 °C with a primary antibody against CD2AP (Proteintech, Wuhan, China).

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: The sections were then incubated overnight at 4 °C with a primary antibody against CD2AP (Proteintech, Wuhan, China).

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: The sections were then incubated overnight at 4 °C with a primary antibody against CD2AP (Proteintech, Wuhan, China).

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: The sections were then incubated overnight at 4 °C with a primary antibody against CD2AP (Proteintech, Wuhan, China).

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: The sections were then incubated overnight at 4 °C with a primary antibody against CD2AP (Proteintech, Wuhan, China).

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: The sections were then incubated overnight at 4 °C with a primary antibody against CD2AP (Proteintech, Wuhan, China).

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: The sections were then incubated overnight at 4 °C with a primary antibody against CD2AP (Proteintech, Wuhan, China).

Techniques: Knockdown, CCK-8 Assay, Migration

Journal: Kidney International Reports

Article Title: Dysregulated Anaerobic Glycolysis in Podocytes is Relevant to the Progression of Focal Segmental Glomerulosclerosis

doi: 10.1016/j.ekir.2025.06.022

Figure Lengend Snippet: Sera from patients with FSGS or LDHA silencing affects α-actinin-4 (ACTN4) expression, leading to actin damage in podocytes. (a) Podocyte-associated genes ( ACTN4 , CD2AP , NPHS2 , and SYNPO ) in serum-treated podocytes from healthy individuals and patients with MCD or FSGS ( n = 5 each). ∗ P < 0.05. (b) Representative western blot analysis of LDHA and ACTN4 in cultured podocytes with or without LDHA silencing ( n = 3). (c) Representative images of F-actin staining and coherency of actin fibers in cultured podocytes with or without LDHA silencing ( n = 5). ∗∗∗∗ P < 0.0001. The scale bar equals 50 μm. (d) Representative images of the podocyte migration and quantification of wound closure in cultured podocytes with or without LDHA silencing ( n = 9). ∗∗ P < 0.01. The scale bar equals 20 μm. ; FSGS, focal segmental glomerulosclerosis; LDHA, lactate dehydrogenase A; MCD, minimal change disease.

Article Snippet: The TaqMan probes and primers used were Hk1 (Mm00439344_m1), Gpi1 (Mm01962484_u1), Pfkm (Mm01309576_m1), Gapdh (Mm99999915_g1), Pgk1 (Mm00435617_m1), Pkm (Mm00834102_gH), Pklr (Mm00443090_m1), Ldha (Mm01612132_g1), Actb (Mm02619580_g1) , ACTN4 (Hs00245168_m1), CD2AP (Hs00961458_m1), NPHS1 (Hs00190446_m1), SYNPO (Hs00702468_s1), and PPIA (Hs04194521_s1).

Techniques: Expressing, Western Blot, Cell Culture, Staining, Migration