Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: Analysis and validation of signature genes. (A) Violin plot of characteristic gene expression in normal and OA samples. (B) Line plot of signature gene expression in normal and OA samples. (C) ROC curves to validate the diagnostic accuracy of signature genes. (D) Expression of ERBB2 in the GSE48556 dataset.

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Biomarker Discovery, Gene Expression, Diagnostic Assay, Expressing

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: Analysis of ERBB2 expression. (A) Heat maps of the top 20 DEGs in high and low ERBB2 expression groups. (B) Volcano map of DEGs in ERBB2 high and low expression groups. (D) Heat map of the correlation between ERBB2 and DEGs, with positive correlation in red and negative correlation in blue.

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Expressing

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: Immune-related function analysis in samples of high and low ERBB2 expression groups (* p < 0.05, ** p < 0.01).

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Expressing

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: GSEA and GSVA analysis of ERBB2. (A–B) GSEA analysis of ERBB2 in high expression group. (C–D) GSEA analysis of ERBB2 in low expression group. (E) E GSVA analysis of biological processes, cellular components, and molecular functions of ERBB2. (F) Pathway-related GSVA analysis of ERBB2.

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Expressing

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: ceRNA Regulatory network and potential drug action network. (A) ceRNA network regulation diagram of lncRNA-miRNA-ERBB2. (B) Potential drug-signature gene network diagram of OA, red for ERBB2, blue for drug.

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques:

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: ERBB2 Transcription levels and protein expression levels between normal and OA samples. (A) Expression level of ERBB2 in qRT-PCR. (B) Protein expression level of ERBB2. (C) Immunohistochemical results of normal human cartilage and OA cartilage from patients (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemical staining

Journal: Annals of Medicine

Article Title: Multi-omics analysis and validation of autophagy-related diagnostic biomarker in osteoarthritis

doi: 10.1080/07853890.2025.2548045

Figure Lengend Snippet: Tissue staining and immunohistochemical analysis. (A) Hematoxylin-eosin staining of normal and OA rat knee joints. (B) so/FG staining of normal and OA rat knee joints. (C) OARSI score. (D-E) Protein expression level and quantitative analysis of ERBB2 in knee cartilage by IHC (* p < 0.05; ** p < 0.01; *** p < 0.001).

Article Snippet: Next, they were blocked with 10% goat serum (Gibco, USA) at room temperature for 20 min and then incubated overnight with the primary antibody of ERBB2 (Proteintech, 18299-1-AP, 1:300).

Techniques: Staining, Immunohistochemical staining, Expressing, Paraffin-embedded Immunohistochemistry

Journal: Cancer letters

Article Title: Isthmin-1 promotes growth and progression of colorectal cancer through the interaction with EGFR and YBX-1.

doi: 10.1016/j.canlet.2024.216868

Figure Lengend Snippet: Fig. 7. Hypoxia elevates ISM1 expression in colon cancer. (A) Colon cancer gene expression data from the TCGA database were grouped by ISM1 expression to perform GSEA analysis. (B) Representative images of HE staining and IHC staining for ISM1 and HIF-1α expression in human colon cancerous tissues. Scale bar = 100 μm. (C) Western blot assay of ISM1 and HIF-1α protein levels in HT-29 cells under physical hypoxia for different period. Densitometry was performed for quanti fication (Right). (D) Effect of HIF-1α inhibitor (BAY87-2243, 10 μM) on ISM1 expression in HT-29 cells under hypoxia conditions. Densitometry was performed for quantification (Right). (E) Plasmids were used to overexpress HIF-1α (OE-HIF-1α) to observe the change of ISM1 expression. Densitometry was performed for quantification (Right). (F) The transcription binding sites of HIF-1α in the promoter region of ISM1 and the positions of the corresponding primers. (G) Dual luciferase reporter gene assay was used to evaluate the effect of HIF-1α on the ISM1 promoter. (H) Enrichment of HIF-1α on the ISM1 promoter as detected by ChIP under hypoxia. (I) The concentration of ISM1 in condition medium (CM) of HT-29 cells under physical hypoxia for different period by using the ELISA kit (n = 3). (J) HT-29 cells without hypoxia were stimulated with the conditional medium (CM) of HT-29 cells after 24 h hypoxia, and then the cells were collected at the following timepoints. EGFR phosphorylation level was detected by western blot assay. Densitometry was performed for quantification (Right). *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: Antibodies against YBX1 (20339- 1-AP), EGFR (66455-1-Ig), HIF-1α (20960-1-AP), GAPDH (HRP-60004) and Tubulin (HRP-66031) were from Proteintech.

Techniques: Expressing, Gene Expression, Staining, Immunohistochemistry, Western Blot, Binding Assay, Luciferase, Reporter Gene Assay, Concentration Assay, Enzyme-linked Immunosorbent Assay, Phospho-proteomics

Journal: Cancer letters

Article Title: Isthmin-1 promotes growth and progression of colorectal cancer through the interaction with EGFR and YBX-1.

doi: 10.1016/j.canlet.2024.216868

Figure Lengend Snippet: Fig. 8. Schematic depiction of ISM1 is transcriptionally regulated by HIF-1α and acts as a novel ligand of EGFR, promoting the progression and metastasis of colorectal cancer via the activation of EGFR signaling pathway.

Article Snippet: Antibodies against YBX1 (20339- 1-AP), EGFR (66455-1-Ig), HIF-1α (20960-1-AP), GAPDH (HRP-60004) and Tubulin (HRP-66031) were from Proteintech.

Techniques: Activation Assay

Journal: Cell reports

Article Title: Granulocyte colony stimulating factor promotes scarless tissue regeneration

doi: 10.1016/j.celrep.2024.114742

Figure Lengend Snippet: (A) Parabiosis between WT:WT ( n = 4, red solid line), CXCR2 KO:CXCR2 KO ( n = 3, blue dotted line), and WT:CXCR2 KO mice ( n = 4, black dotted line). Shown is the percentage of ear hole closure. 2-way ANOVA comparing WT:KO pairs to WT:WT pairs. (B) ELISA measuring cytokine expression in injured WT ( n = 4 for day 3, n = 3 for day 7) and CXCR2 KO ( n = 6 for day 3, n = 3 for day 7) plasma. Unpaired two-tailed Student’s t test. (C) WT ( n = 6), CXCR2 KO ( n = 7) and G-CSF depleted CXCR2 KO ( n = 4) plasma was injected daily into the wound bed of WT mice undergoing WIHN for the first 3 days after injury. Shown are representative photographs and quantification of hair follicles. Scale bars, 100 μM. Unpaired two-tailed Student’s t test. (D) G-CSF ( n = 9) or PBS (control, n = 7 ) was injected daily into the wound bed of WT mice undergoing WIHN for the first 3 days after injury. Representative photographs of whole-mount and scanning electron microscopy demonstrating unpigmented hairs in the center of the healed areas. Right: quantification of hair follicles. Scale bars, 100 μM. Unpaired two-tailed Student’s t test. (E) Representative immunofluorescence images of PBS and G-CSF-injected WT wound beds depicting hair follicle structures (Krt14+, Krt6+) Scale bars, 100 μM. (F) Representative photographs and quantification of scar size of G-CSF-treated ( n = 8) or PBS-treated (control, n = 3 ) stented back wounds at day 28 after injury. Scale bars, 1 mm. Unpaired two-tailed Student’s t test. (G) Representative trichrome-stained tissue sections from G-CSF- or PBS-treated stented back wounds. A black line highlights scar size. (H) Quantification of scar diameter for G-CSF ( n = 6) or PBS-treated ( n = 3) mice. Unpaired two-tailed Student’s t test. (I) Wound fibrosis assessed by picrosirius red staining in G-CSF-treated ( n = 14 sections) or PBS-treated ( n = 10 sections). Unpaired two-tailed Student’s t test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Mean ± SEM are plotted.

Article Snippet: G-CSF , Med Chem Express , HY-P70608.

Techniques: Enzyme-linked Immunosorbent Assay, Expressing, Clinical Proteomics, Two Tailed Test, Injection, Control, Electron Microscopy, Immunofluorescence, Staining

Journal: Cell reports

Article Title: Granulocyte colony stimulating factor promotes scarless tissue regeneration

doi: 10.1016/j.celrep.2024.114742

Figure Lengend Snippet: (A) Dot plot demonstrating average expression and percentage of immune cells expressing Csf3r . (B) Analysis of key cell-to-cell interactions between immune cells in the ear skin of WT (salmon color) and CXCR2 KO (blue color) mice. Mac, macrophage; T, T cell. (C) Dot plot demonstrating average gene expression of key genes between WT and CXCR2 KO macrophages. (D) GSEA of macrophage populations in WT and CXCR2 KO wounded skin. (E) Representative images and quantification of immunofluorescence of WT and CXCR2 KO wounded skin for CD80 ( n = 4), COX2 ( n = 4), CD163 ( n = 12 for WT and n = 11 for KO), MRC1 ( n = 10 for WT and n = 6 for KO), and ARG1 ( n = 10 for WT and n = 6 for KO). Cell percentages are calculated with total DAPI+ cells as the denominator. Scale bars, 100 μM. Unpaired two-tailed Student’s t test. (F) Representative H&E immunostaining images and quantification of PBS- and G-CSF-injected stented back wounds of WT mice for CD163 ( n = 7 for PBS and n = 5 for G-CSF), MRC1 ( n = 5 for PBS and n = 7 for G-CSF), CD31 ( n = 8 for PBS and n = 7 for G-CSF) and Ki67 ( n = 5 for PBS and n = 4 for G-CSF). Cell percentages are calculated with total DAPI+ cells as the denominator. Scale bars, 100 μM. Unpaired two-tailed Student’s t test. * p < 0.05, ** p < 0.01, *** p < 0.01. Mean ± SEM are plotted.

Article Snippet: G-CSF , Med Chem Express , HY-P70608.

Techniques: Expressing, Gene Expression, Immunofluorescence, Two Tailed Test, Immunostaining, Injection