Journal: Frontiers in Genetics

Article Title: Integrative analysis of single-cell and microarray data reveals SPI1-centered macrophage regulatory signatures in ulcerative colitis

doi: 10.3389/fgene.2025.1617834

Figure Lengend Snippet: SPI1(+) mainly regulates UC macrophage activation. (A) GSEA plots showing significant enrichment of macrophage activation-associated pathway in UC. (B) Transcription factors regulating macrophage activation-associated pathway including SPI1 (+). (C) Rank for regulons in macrophages based on the RSS. (D) Macrophages are highlighted in the UMA; (E) Binarized RAS for the top regulon SPI1 on UMAP. (F) UMAP showing SPI1 regulon activity across cell types. (G) Average expression of SPI1 regulon genes in macrophages. Data are expressed as mean ± SD. *P < 0.05, **P < 0 .01 compared to the other group.

Article Snippet: ELISA kit for IL-1β (ZC-37974, Zhuocai, China); ELISA kit for IL-10 (ZC-37962, Zhuocai, China); SPI1 antibody (55100-1-AP, Proteintech, China); iNOS antibody (22226-1-AP, Proteintech, China); Arg1 antibody (16001-1-AP, Proteintech, China); Tubulin antibody (80762-1-RR, Proteintech, China); Anti-rabbit IgG (H + L) (14780, Cell Signaling Technology, United States).

Techniques: Activation Assay, Activity Assay, Expressing

Journal: Frontiers in Genetics

Article Title: Integrative analysis of single-cell and microarray data reveals SPI1-centered macrophage regulatory signatures in ulcerative colitis

doi: 10.3389/fgene.2025.1617834

Figure Lengend Snippet: Identification of SPI1-regulated hub genes through the integration of gene set intersection and machine learning approaches. (A,B) LASSO regression analysis with cross-validation curve showing the optimal lambda value selection and coefficient profiles for gene selection. (C) Accuracy versus number of variables selected using RFE. (D) Bar chart of top genes ranked by importance scores. (E) SVM-RFE identified a 30-gene signature that achieved the highest classification accuracy. (F) Venn diagram illustrating the overlap between genes selected by three machine learning methods (LASSO, RFE-RF, and SVM-RFE). (G,H) Expression levels of SPI1, IRAK3, IL1RN, CD55, and PEA15 in two independent datasets GSE87466 (G) and GSE75214 (H) . (I) ROC curves for IRAK3, IL1RN, CD55 and PEA15 in the training dataset. (J) ROC curves for IRAK3, IL1RN, CD55 and PEA15 in the validation dataset. **** P < 0.0001 compared to the control group.

Article Snippet: ELISA kit for IL-1β (ZC-37974, Zhuocai, China); ELISA kit for IL-10 (ZC-37962, Zhuocai, China); SPI1 antibody (55100-1-AP, Proteintech, China); iNOS antibody (22226-1-AP, Proteintech, China); Arg1 antibody (16001-1-AP, Proteintech, China); Tubulin antibody (80762-1-RR, Proteintech, China); Anti-rabbit IgG (H + L) (14780, Cell Signaling Technology, United States).

Techniques: Biomarker Discovery, Selection, Expressing, Control

Journal: Frontiers in Genetics

Article Title: Integrative analysis of single-cell and microarray data reveals SPI1-centered macrophage regulatory signatures in ulcerative colitis

doi: 10.3389/fgene.2025.1617834

Figure Lengend Snippet: LPS-induced polarization of M1 macrophages is affected by SPI1 in vitro. (A–D) The protein expressions of SPI1 (B) , iNOS (C) , Arg1 (D) measured by WB. (E–G) Macrophage M1 polarization induced by LPS was detected by flow cytometry. (H, I) WB analysis was performed to assess the knockdown efficiency of SPI1 following si-SPI1 treatment, including three groups: untreated control, si-NC (non-targeting siRNA), and si-SPI1. (J–L) The expressions of iNOS (K) and Arg1 (L) measured by WB after SPI1 knockdown. (M–P) Macrophage polarization induced by LPS was detected by flow cytometry after SPI1 knockdown. (Q, R) Levels of IL-10 (Q) and IL-1β (R) measured by ELISA after SPI1 knockdown. All experiments were performed in triplicate with independently prepared biological samples. Data are expressed as mean ± SD. * P < 0.05, ** P < 0 .01 compared to the other group.

Article Snippet: ELISA kit for IL-1β (ZC-37974, Zhuocai, China); ELISA kit for IL-10 (ZC-37962, Zhuocai, China); SPI1 antibody (55100-1-AP, Proteintech, China); iNOS antibody (22226-1-AP, Proteintech, China); Arg1 antibody (16001-1-AP, Proteintech, China); Tubulin antibody (80762-1-RR, Proteintech, China); Anti-rabbit IgG (H + L) (14780, Cell Signaling Technology, United States).

Techniques: In Vitro, Flow Cytometry, Knockdown, Control, Enzyme-linked Immunosorbent Assay