Journal: Frontiers in Immunology
Article Title: Meta-analysis of multi-center transcriptomic profiles and machine learning reveal phospholipase Cβ4 as a Wnt/Ca² + signaling mediator in glioblastoma immunotherapy
doi: 10.3389/fimmu.2025.1610683
Figure Lengend Snippet: In vitro experiments showing that PLCB4 in the ProImmuML signature may be a downstream regulator of the Wnt pathway and inhibit GBM proliferation. (A) Metascape analysis of the ProImmuML signature. (B) qPCR analysis was performed to analyze the effects of three different Wnt pathway agonists/inhibitors on the expression of APCDD1 (left panel), RAC2 (middle panel), and PLCB4 (right panel). SKL: SKL-2001, Wnt/β-catenin signaling pathway agonist; MSAB: MSAB, Wnt/β-catenin signaling pathway inhibitor; 2-APB: 2-APB, Wnt/Ca 2+ signaling pathway inhibitor; LON: Lonomycin, Wnt/Ca 2+ signaling pathway agonist; BLE: Blebbistatin, Planar cell polarity pathway inhibitor; WNT: Wnt5a, Planar cell polarity pathway agonist. (C) Representative IHC staining images of PLCB4 from four glioma patients with higher expression of PLCB4 (left panel, n = 2) and lower expression of PLCB4 (right panel, n = 2). (D) The intersection of genes from two parallel comparisons identified 235 upregulated genes (left panel) and 65 downregulated genes (right panel). (E) KEGG enrichment analysis of the upregulated genes. (F) GSEA revealing proliferation-related pathways were significantly inhibited after overexpressing PLCB4. (G) EdU assay showing PLCB4 inhibiting the proliferation of glioma cells in U87. Significant difference, *P<0.05, **P<0.01, ***P<0.001.
Article Snippet: Lonomycin (LON; TargetMol, China) and 2-APB (TargetMol, China) served as the activator and inhibitor of the Wnt/Ca2 + pathway, while Wnt5a (TargetMol, China) and blebbistatin (TargetMol, China) were used to modulate the Wnt/PCP pathway.
Techniques: In Vitro, Expressing, Immunohistochemistry, EdU Assay
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