Fig 1: expression profile of proteins involved in certain aspects of cancer development such as cell proliferation, glycolytic metabolism, and cell invasion. (A) Total protein extracts from control and transduced HaCaT cells were obtained using a specific lysis buffer (Lysis buffer 17) (R&D Systems, Catalogue # 895943) included in the array kit. The relative expression levels of three proteins involved in cell proliferation and metabolism pathways (FGF basic, Enolase 2, and Fox01/FKHR) in HaCaT transduced and co-transduced cells compared to that in control cells is shown. (B) Relative expression levels of proteins expressed as mean spot pixel density that are involved in cell invasion (uPA, and CapG) and one protein involved in oxidative stress responses (HO-1/HMOX1) are shown for HaCaT transduced and co-transduced cells compared to those of control cells as assessed by the Human XL Oncology Array Kit (R&D Systems, MN, USA, ARY026). Averages ± standard deviations (SDs) of two independent experiments are shown. A two-way analysis of variance (ANOVA) test was conducted to assess statistical significance among groups along with Dunnett post-hoc test for intra-group comparison. (*) p < 0.05, (**) p < 0.01, (***) p < 0.001.
Fig 2: Comparison of 9 cell lines in the form of single cells (left) and tumorospheres (TOS) (right) regarding the expression of 43 of the 85 cancer-related proteins detected by a Proteome Profiler Human XL Oncology Array (R&D Systems) (Catalog # ARY026).
Fig 3: Proteome profiling of oncology-related markers in MDA-MB-231 cells treated with wild ginseng adventitious root extract (WGAR) and cisplatin. Cells were treated with WGAR or cisplatin at the 20% inhibitory concentration (IC20; 3 μg/mL for WGAR, 2 μg/mL for cisplatin) and 50% inhibitory concentration (IC50; 79 μg/mL for WGAR, 9 μg/mL for cisplatin) for 48 h. Cell lysates (200 μg) were analyzed using the Proteome Profiler Human XL Oncology Array (R&D Systems, ARY026). (a) Representative array membrane images. (b) Quantification of selected oncology-related markers at IC20 concentrations. Proteins are grouped by functional categories as indicated by horizontal lines below the x-axis. (c) Quantification of selected oncology-related markers at IC50 concentrations. Proteins are grouped by functional categories as indicated by horizontal lines below the x-axis. (d) Independent validation of IL-6 and M-CSF by ELISA. IL-6 and M-CSF levels in culture supernatants were quantified using the Human IL-6 Quantikine ELISA Kit (Cat# D6050, R&D Systems) and Human M-CSF Quantikine ELISA Kit (Cat# DMC00B, R&D Systems), respectively, and expressed as fold change relative to control (set to 1.0). Data are expressed as fold change relative to control (dashed line = 1). Values represent mean ± SD from three independent experiments. Statistical significance was determined by one-way ANOVA followed by Dunnett’s post hoc test: * p < 0.05, ** p < 0.01, *** p < 0.001 compared to the untreated control. CCL2, C-C motif chemokine ligand 2; CXCL8, C-X-C motif chemokine ligand 8; Dkk-1, Dickkopf-1; GM-CSF, granulocyte-macrophage colony-stimulating factor; IL-6, interleukin-6; M-CSF, macrophage colony-stimulating factor; PAI-1, plasminogen activator inhibitor-1; u-PA, urokinase-type plasminogen activator; VEGF, vascular endothelial growth factor; WGAR, wild ginseng adventitious root.
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