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SDF4 influences the cell cycle progression and apoptosis in glioma cells. A , B Representative Annexin V–FITC/PI flow-cytometry plots and quantification showing increased apoptosis after SDF4 knockdown in H4 and SW1783 cells. C , D Cell-cycle analysis showing an increased G0/G1 fraction and reduced S-phase fraction, indicating G0/G1 arrest. E Immunoblotting showed increased Bax and cleaved caspase-3 and decreased Bcl-2 after SDF4 knockdown. F SDF4 knockdown reduced Cyclin D1, CDK4, and CDK6 and increased p21 and p27. G KEGG enrichment analysis identified PI3K-AKT signaling as a candidate pathway for validation. H SDF4 knockdown reduced p-PI3K and p-AKT levels, whereas total PI3K and AKT were comparatively unchanged, supporting an association between SDF4 and PI3K-AKT phosphorylation rather than a direct activation mechanism. Flow-cytometry data are presented as mean ± SD from independent experiments and were analyzed using the two-group test specified in Methods. * P < 0.05. Immunoblots are representative of independent experiments. FITC, fluorescein isothiocyanate; PI, propidium iodide; KEGG, Kyoto Encyclopedia of Genes and Genomes; p-PI3K, phosphorylated PI3K; p-AKT, phosphorylated AKT

Journal: BMC Cancer

Article Title: Integrative transcriptomic and experimental analyses identifies SDF4 as an astrocyte-expressed regulator of glioma progression through PI3K-AKT signaling

doi: 10.1186/s12885-026-16635-6

Figure Lengend Snippet: SDF4 influences the cell cycle progression and apoptosis in glioma cells. A , B Representative Annexin V–FITC/PI flow-cytometry plots and quantification showing increased apoptosis after SDF4 knockdown in H4 and SW1783 cells. C , D Cell-cycle analysis showing an increased G0/G1 fraction and reduced S-phase fraction, indicating G0/G1 arrest. E Immunoblotting showed increased Bax and cleaved caspase-3 and decreased Bcl-2 after SDF4 knockdown. F SDF4 knockdown reduced Cyclin D1, CDK4, and CDK6 and increased p21 and p27. G KEGG enrichment analysis identified PI3K-AKT signaling as a candidate pathway for validation. H SDF4 knockdown reduced p-PI3K and p-AKT levels, whereas total PI3K and AKT were comparatively unchanged, supporting an association between SDF4 and PI3K-AKT phosphorylation rather than a direct activation mechanism. Flow-cytometry data are presented as mean ± SD from independent experiments and were analyzed using the two-group test specified in Methods. * P < 0.05. Immunoblots are representative of independent experiments. FITC, fluorescein isothiocyanate; PI, propidium iodide; KEGG, Kyoto Encyclopedia of Genes and Genomes; p-PI3K, phosphorylated PI3K; p-AKT, phosphorylated AKT

Article Snippet: The antibodies included Cyclin D1 (dilution 1:3000; Abcam), CDK4 (dilution 1:1000; CST), CDK6 (dilution 1:1000; Proteintech), P21 (dilution 1:1000; CST), P27 (dilution 1:1000; CST), BAX (dilution 1:2000; CST), Bcl-2 (dilution 1:1000; Abcam), Cleaved caspase-3 (dilution 1:500; Affbiotech), SDF4 (dilution 1:500; Santa Cruz Biotechnology), p-PI3K (dilution 1:1000; Affbiotech), PI3K (dilution 1:2000; CST), p-AKT (dilution 1:1000; CST), AKT (dilution 1:2000; CST), and β-actin (dilution 1:10000; Beijing TDY Biotechnology Co.) targeting various proteins of interest.

Techniques: Flow Cytometry, Knockdown, Cell Cycle Assay, Western Blot, Analysis, Biomarker Discovery, Phospho-proteomics, Activation Assay