Journal: Journal of Biological Chemistry

Article Title: Silencing of Insulin-like Growth Factor-binding Protein-2 in Human Glioblastoma Cells Reduces Both Invasiveness and Expression of Progression-associated Gene CD24

doi: 10.1074/jbc.m609567200

Figure Lengend Snippet: FIGURE 2. Modulation of IGF/IGF-IR signaling caused by knockdown of IGFBP-2. The IGFBP-2-knockdown cells (IGFBP-2 KD) and control cells were treated with the indicated concentrations of IGF-I or IGF-II in serum-free con- dition, and extents of phosphorylation of IGF-IR (p-IGF-IR), Akt (p-Akt), and ERK1/2 (p-Erk1/2) were analyzed by immunoblot and a representative result of U251 was shown. The band intensity was measured and the ratio of phos- phorylated protein to the corresponding total protein was calculated.

Article Snippet: The following primary antibodies were used: anti-human IGFBP-2 mouse monoclonal antibody (C-10; Santa Cruz Biotechnology, Santa Cruz, CA), anti- -actinmousemonoclonal antibody (AC-74; Sigma), anti-phosphoERK1/2 rabbit monoclonal antibody (Thr185/Tyr187, clone AW39;Upstate Cell Signaling Solutions, Lake Placid, NY), antiphospho-IGF-IR rabbit polyclonal antibody (Tyr1131), antiIGF-IR rabbit polyclonal antibody, anti-phospho-Akt mouse monoclonal antibody (Ser473), anti-Akt rabbit polyclonal antibody, anti-phospho-ERK1/2 rabbit monoclonal antibody (Thr185/Tyr187, clone AW39; Upstate Cell Signaling Solutions, Lake Placid, NY), anti-ERK1/2 rabbit polyclonal antibody, anticaspase-3, and cleaved capase-3 (Asp175) rabbit polyclonal antibodies, and anti-heat shock protein 70 rabbit polyclonal antibody (Cell Signaling Technology, Danvers, MA).

Techniques: Knockdown, Control, Phospho-proteomics, Western Blot

Journal: Journal of Inflammation Research

Article Title: Single-Cell Sequencing and Machine Learning Integration to Identify Candidate Biomarkers in Psoriasis: INSIG1

doi: 10.2147/JIR.S492875

Figure Lengend Snippet: Advanced multi-machine learning analyses identify INSIG1 as an emerging biomarker for psoriasis. ( A ) DEGs in the GSE14905 and GSE66511 dataset. Red circles represent upregulated genes while blue circles represent downregulated genes. ( B ) The DEGs that are commonly upregulated and downregulated in the GSE14905 and GSE66511 datasets. ( C and D ) Results of GO and KEGG enrichment analysis of the new 110 DEGs. ( E ) Visualization of interactive genes from three machine learning methods. ( F ) ROC curve reflecting the diagnostic performance of INSIG1 across GSE14905 and GSE66511 datasets and the expression of INSIG1 in the Pso and Control groups of the two datasets. ( G ) Box plots of differential expression for INSIG1 in the GSE14905 and GSE66511 datasets. ( H ) Bar charts illustrating the expression levels of INSIG1 in T cells between individuals with psoriasis and those from a healthy cohort within the scRNA-seq dataset of GSE62183.

Article Snippet: Endogenous peroxidase activity was inhibited, followed by an overnight incubation at 4°C with an anti- INSIG1 antibody (Proteintech, 55282-1-AP).

Techniques: Biomarker Discovery, Diagnostic Assay, Expressing, Control, Quantitative Proteomics

Journal: Journal of Inflammation Research

Article Title: Single-Cell Sequencing and Machine Learning Integration to Identify Candidate Biomarkers in Psoriasis: INSIG1

doi: 10.2147/JIR.S492875

Figure Lengend Snippet: Functional enrichment and Spearman correlation analysis of INSIG1 co-expressed genes. ( A ) GO enrichment analysis of INSIG1 co-expressed genes. ( B ) KEGG enrichment analysis of INSIG1 co-expressed genes. ( C ) Results of correlation between INSIG1 expression and T cell subtypes via Spearman correlation analysis.

Article Snippet: Endogenous peroxidase activity was inhibited, followed by an overnight incubation at 4°C with an anti- INSIG1 antibody (Proteintech, 55282-1-AP).

Techniques: Functional Assay, Expressing

Journal: Journal of Inflammation Research

Article Title: Single-Cell Sequencing and Machine Learning Integration to Identify Candidate Biomarkers in Psoriasis: INSIG1

doi: 10.2147/JIR.S492875

Figure Lengend Snippet: Cellular communication analysis and ligand–receptor interactions of INSIG1 expressing T cells and other cells. ( A ) Expression level of high and low INSIG1 expressing T cells. ( B ) Cell communication analysis between high and low INSIG1 expressing T cells and other cells. ( C and D ) Ligand–receptor interactions between high and low INSIG1 expressing T cells and other cells.

Article Snippet: Endogenous peroxidase activity was inhibited, followed by an overnight incubation at 4°C with an anti- INSIG1 antibody (Proteintech, 55282-1-AP).

Techniques: Expressing

Journal: Journal of Inflammation Research

Article Title: Single-Cell Sequencing and Machine Learning Integration to Identify Candidate Biomarkers in Psoriasis: INSIG1

doi: 10.2147/JIR.S492875

Figure Lengend Snippet: Validation of INSIG1 expression in psoriasis through experiments and molecular docking results between INSIG1 and tetrandrine. ( A and B ) IHC results of INSIG1 expression in the Pso group compared with the Control group in clinical samples (n=20) and ( C and D ) in in vivo (n=6) experiments (scale bar = 50 μm and 20 μm). ( E ) mRNA expression level of INSIG1 in the Pso model group compared with the control group in vivo (n=6). ( F ) Drug sensitivity enrichment analysis of INSIG1 . ( G and H ) Visualization of molecular docking between INSIG1 and tetrandrine. ( I ) Results of interaction analysis between ligand (tetrandrine) and receptor ( INSIG1 ). Statistical significance: *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001, compared with the Control group.

Article Snippet: Endogenous peroxidase activity was inhibited, followed by an overnight incubation at 4°C with an anti- INSIG1 antibody (Proteintech, 55282-1-AP).

Techniques: Biomarker Discovery, Expressing, Control, In Vivo

Journal: iScience

Article Title: FAM65A, as a potential predictor of prognosis, promotes colorectal cancer progression via activating Ras/ERK/RSK signaling

doi: 10.1016/j.isci.2026.114662

Figure Lengend Snippet: FAM65A binds to Ras and activates the Ras/ERK signaling to mediate RSK activation (A) The volcano plot analysis results for the FAM65A high-expression and low-expression groups from the TCGA database were shown. (B) The KEGG and GO results were shown. (C) The GSEA results were shown. (D) GSEA on DEGs between the FAM65A high-expression group and low-expression group in the Reactome database were shown. (E) IP was performed to detect the binding of FAM65A and Ras/p-RSK. (F) IP was performed to detect the binding of Ras and FAM65A/p-RSK. (G) Immunofluorescence was performed to detect the co-localization of FAM65A and Ras. Scale bars, 20 μm. (H) Western blot analysis the Ras and p -ERK expression in FAM65A knockdown or overexpression cells. Data are presented as mean ± SEM of biologically independent experiments.

Article Snippet: p -ERK , Proteintech , Cat# 28733-1-AP; RRID: AB_2881202.

Techniques: Activation Assay, Expressing, Binding Assay, Immunofluorescence, Western Blot, Knockdown, Over Expression

Journal: iScience

Article Title: FAM65A, as a potential predictor of prognosis, promotes colorectal cancer progression via activating Ras/ERK/RSK signaling

doi: 10.1016/j.isci.2026.114662

Figure Lengend Snippet: Ras/ERK signaling activation was indispensable for FAM65A-mediated RSK activation and CRC progression (A) Western blot analysis of Ras and p -ERK expression in HCT116-FAM65A cells treated with 10 μM Abd-7, or without treatment. (B) Results from the CCK8 cell proliferation assay conducted on HCT116-FAM65A cells with and without the application of Abd-7, n = 3, ∗∗∗ p < 0.001. (C) Colony formation assay performed on HCT116-FAM65A cells treated with Abd-7 or not. (D) Quantitative analysis of the colony formation assay results, n = 3, ∗∗∗ p < 0.001. (E) Results from the EdU assay conducted on HCT116-FAM65A cells with and without the application of Abd-7. Scale bars, 100 μm. (F) Quantitative analysis of the EdU assay results, n = 3, ∗∗∗ p < 0.001. (G) Western blot analysis of Ki-67, cleaved Caspase 3, Bcl-2, and Bax expression in HCT116-FAM65A cells treated with Abd-7 or not. (H) Results from the apoptosis assay conducted on HCT116-FAM65A cells treated with Abd-7 or not. Scale bars, 50 μm. (I) Quantitative analysis of the apoptosis experiments, n = 3, ∗∗∗ p < 0.001. (J) Results from the Transwell migration assay conducted on HCT116-FAM65A cells with and without the application of Abd-7. Scale bars, 50 μm. (K) Quantitative analysis of the Transwell migration assay results, n = 3, ∗∗∗ p < 0.001. (L) Results from the wound healing assay performed on HCT116-FAM65A cells treated with Abd-7 or not. Scale bars, 50 μm. (M) Quantitative analysis of the wound healing assay results, n = 3, ∗∗∗ p < 0.001. (N) Western blot analysis the expression of EMT markers in HCT116-FAM65A cells treated with Abd-7 or not. (O) Proposed model of FAM65A in CRC progression. Data are presented as mean ± SEM of biologically independent experiments.

Article Snippet: p -ERK , Proteintech , Cat# 28733-1-AP; RRID: AB_2881202.

Techniques: Activation Assay, Western Blot, Expressing, Proliferation Assay, Colony Assay, EdU Assay, Apoptosis Assay, Transwell Migration Assay, Wound Healing Assay

Journal: iScience

Article Title: FAM65A, as a potential predictor of prognosis, promotes colorectal cancer progression via activating Ras/ERK/RSK signaling

doi: 10.1016/j.isci.2026.114662

Figure Lengend Snippet: Knockdown of FAM65A inhibits tumor progression in vivo (A) LOVO-shCtrl and LOVO-shFAM65A cells were administered into the fourth fat pad of nude mice, and the resulting tumor growth curves were subsequently generated, n = 5, ∗ p < 0.05. (B) The tumors excised from mice across various experimental groups are presented. (C) Hematoxylin and Eosin (HE) staining results of lung tissue from the different groups is displayed. (D) A quantitative analysis of metastatic lung nodules is provided, n = 5, ∗∗ p < 0.01. (E) IHC results for FAM65A, Ki-67, p -RSK, p -ERK, Ras, N-cadherin, vimentin, cleaved Caspase 3, ZO-1, and E-cadherin in tumor tissues are illustrated. (F) A quantitative analysis of the IHC results is included. Data are presented as mean ± SEM of biologically independent experiments, n = 5, ∗∗∗ p < 0.001.

Article Snippet: p -ERK , Proteintech , Cat# 28733-1-AP; RRID: AB_2881202.

Techniques: Knockdown, In Vivo, Generated, Staining