Fig 1: DPMSC-CM cytokine profile after 48 h incubation with DMEM + 0.2% FBS (n = 5). IL-4: Interleukin 4, IL-2: Interleukin 2, CXCL10: C-X-C motif chemokine ligand 10, IL-1β: Interleukin 1 beta, TNF-α: Tumor necrosis factor alpha, CCL2: C-C motif chemokine ligand 2, IL-17A: Interleukin 17A, IL-6: Interleukin 6, IL-10: Interleukin 10, IFN-γ: Interferon gamma, IL-12p70: Interleukin 12, CXCL8: Interleukin 8, TGF-β1: Transforming growth factor beta 1.
Fig 2: Stt + Tcz decreases pSTAT-3 at 24 h in DU-145 cells. (A) Effect of Stt on DU-145 viability was determined at 1, 3, 5, 7 and 10 µM at 24 h. (B) Effect of Tcz was determined at 10, 100, 1,000 and 10,000 ng/ml. (C) In-Cell Western assay was used to evaluate the effect of Stt, Tcz, IL-6 for 24 h. (D) Densitometric analysis. Data are presented as the mean ± SD of three independent experiments, normalized to their respective control. *P<0.05, **P<0.01 and ***P<0.001 vs. UCG. Stt, stattic; Tcz, tocilizumab; p, phosphorylated; Et, Etoposide; UCG, untreated control group; OD, optical density.
Fig 3: Stt + Tcz decreases VEGF and vimentin and increases E-cadherin expression induced by IL-6 in DU-145 cells. (A) VEGF in DU-145 cell supernatant was assessed by ELISA. (B) Western blotting was utilized to evaluate the differences in E-cadherin and vimentin expression. Densitometric analysis of (C) vimentin and (D) E-cadherin. Data are presented as the mean ± SD of three independent experiments. *P<0.05, **P<0.01 and ***P<0.001. Stt, stattic; Tcz, tocilizumab.
Fig 4: Stt + Tcz treatment decreases migratory and invasive capacity induced by IL-6 in the DU-145 prostate cancer cells. (A) Representative wound-healing assay. Scale bar, 100 µm (photographs were obtained with 40X amplification), (B) closure rate and (C) effect of different treatment at 24 h. (D) Chemotaxis assay was performed to evaluate invasive cells, the scale bars denote 40 µm (100× amplification) and (E) SRB was measured. Data are presented as the mean ± SD of three independent experiments, normalized to respective UCG. *P<0.05, **P<0.01 and ***P<0.001. Stt, stattic; Tcz, tocilizumab; UCG, untreated control group; OD, optical density; SRB, Sulforhodamine B.
Fig 5: DPMSC-S induced significant alterations in the growth factor, pro-inflammatory and anti-inflammatory cytokine levels of AW13516, MDA-MB-231, and A375 cell lines: AW13516, MDA-MB-231, and A375 cell lines were treated with 20%, 50%, and 100% DPMSC-CM for 48 h, and conditioned media from these cells were collected post 48 h of treatment to perform flow cytometry-based growth factor analysis in the secretome for Angiopoietin-2, EGF, EPO, FGF basic, G-CSF, GM-CSF, HGF, M−CSF, PDGF-AA, PDGF-BB, SCF, TGF-α, VEGF. (a) Comparative analysis of growth factors in AW13516 cell line secretome. (b) Comparative analysis of growth factors in MDA-MB-231 cell line secretome. (c) Comparative analysis of growth factors in A375 cell line secretome. AW13516, MDA-MB-231, and A375 cell lines were treated with 20%, 50%, and 100% DPMSC-CM for 48 h, and conditioned media from these cells were collected post 48 h of treatment to perform flow cytometry-based cytokine analysis in the secretome for IL-4, IL-2, CXCL10, IL-1β, TNF-α, CCL2, IL-17A, IL-6, IL-10, IFN-γ, IL-12p70, CXCL8, TGF-β1. (d) Comparative analysis of cytokines in AW13516 cell line secretome. (e) Comparative analysis of cytokines in MDA-MB-231 cell line secretome. (f) Comparative analysis of cytokines in A375 cell line secretome. Experiments were performed in triplicates and repeated 2 times. ns not significant, *p < 0.05, **p < 0.01.
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