PSPH Antibody - #DF12711

Fig. 4: CXCL7 upregulates PHGDH and activates serine metabolism in cancer cells. A Left: GSEA of enriched pathways in differentially expressed genes (DEGs) between HT29-control and HT29-CXCL7 groups. Right: Comparative analysis of PHGDH FPKM values between groups. B The expression level of PHGDH in control and CXCL7-overexpressing cells with or without the CXCR2 inhibitor, as determined by RT-qPCR. C Western blot detection of serine biosynthesis enzymes (PHGDH, PSAT1, PSPH).GAPDH was used as the loading control. D Kaplan‒Meier analysis of overall survival in TCGA-CRC patients stratified by PHGDH expression. E Left: Metabolite enrichment pathway bubble plot. Middle: Heatmap of differential metabolites. Right: Schematic of serine/one-carbon metabolism. F Heatmap visualization of DEGs in glycine, serine, and threonine metabolism pathways. G ELISA analysis of SAM levels in CXCL7-overexpressing cells and PHGDH-inhibited cells. H TUNEL assays of CXCL7-overexpressing cells transfected with NC-siRNA or PHGDH-siRNA, with or without SAM treatment(scale bar, 50 μm). I The drug sensitivity of CXCL7-overexpressing cells transfected with NC-siRNA or PHGDH-siRNA, with or without SAM treatment. All data are presented as mean ± SD. *P 

Figure 2. PHGDH is involved in the cellular senescence of hDPCs. (a) Analysis of the transcription of PHGDH, PSAT1, and PSPH in different passages (P5 and P12) of hDPCs by RT-qPCR. (b) Analysis of PHGDH, PSAT1, and PSPH expression in different passages (P5 and P12) of hDPCs by western blot. (c) Young hDPCs (P5) were treated with different concentrations of NCT—503 and CBR—5884 for 48 h, and cell viability was determined by CCK8 assay. (d) Effects of NCT—503 (20 μM) and CBR—5884 (30 μM) treatment for 48 h on young hDPCs (P5) were determined by SA-β-gal staining, Ki67 staining, and yH2AX staining. In SA-β-gal staining, blue-stained cells are positive cells. In Ki67 staining and yH2AX staining, the blue color in the nucleus shows DAPI staining, the green color in the nucleus shows Ki67 or yH2AX staining. (e) Transcription of LMNB1 and P21 in hDPCs (P5) treated with NCT—503 (20 μM) and CBR—5884 (30 μM) for 48 h was analyzed by RT-qPCR. (f) Crystal violet staining determined colony formation at 7 days after continuous treatment with NCT—503 (20 μM) and 2 days of CBR—5884 (30 μM) treatment. Data were expressed as means ± SD, n = 3 independent experiments. * p < 0.05.