Fig 1: Effect of aminoguanidine on IGF-1-induced MG-derived glycative stress in LNCaP cells. LNCaP cells were pretreated with aminoguanidine (AG, 1 mM) for 6 h before exposure to IGF-1 (150 ng/mL) for 72 h. MG-derived glycative stress was evaluated by measuring (a) intracellular MG-H1 levels using a specific ELISA assay; GLO1 expression at the (b) mRNA level by qRT-PCR and (c) protein level by Western blot analysis; and (d) GLO1 enzymatic activity using a spectrophotometric assay. β-Actin was used as the loading control for normalization of Western blot data. Data are presented as the mean ± SD of three independent experiments; each black dot represents one biological replicate. *** p < 0.001; **** p < 0.0001.
Fig 2: Effect of IGF-1 on RAGE and OPN mRNA expression in LNCaP cells. LNCaP cells were treated with IGF-1 (150 ng/mL) for 72 h or left untreated (control). (a) RAGE and (b) OPN mRNA expression was evaluated by quantitative real-time PCR (qRT-PCR). Data are presented as the mean ± standard deviation (SD) of three independent experiments; each black dot represents one biological replicate. ** p < 0.01, *** p < 0.001 versus untreated cells.
Fig 3: Effect of aminoguanidine on IGF-1-induced cell proliferation and invasive capacity in LNCaP cells. LNCaP cells were pretreated with aminoguanidine (AG, 1 mM) for 6 h before exposure to IGF-1 (150 ng/mL) for 72 h. Cell proliferation was evaluated by (a) cell counting and (b) colony formation assay. Invasive capacity was assessed by (c) a cell invasion assay and by evaluating the mRNA expression of the invasion-related metalloproteinases (d) MMP-1, MMP-7, and MMP-9 using qRT-PCR. Data are presented as the mean ± SD of three independent experiments; each black dot represents one biological replicate. * p < 0.05; ** p < 0.01; *** p < 0.001, **** p < 0.0001.
Fig 4: Basal IGF-1 expression is associated with increased MG-derived glycative stress in aggressive prostate cancer (PCa) cells. LNCaP and PC3 cells, representing less aggressive, androgen-dependent and more aggressive, androgen-independent human prostate cancer (PCa) cell models, respectively, were analyzed for IGF-1 expression at the (a) mRNA level by quantitative real-time PCR (qRT-PCR) and (b) intracellular protein level by enzyme-linked immunosorbent assay (ELISA), (c) MG-H1 levels by ELISA, and (d) GLO1 specific activity by spectrophotometric assay. Data are presented as the mean ± standard deviation (SD) of three independent experiments; each black dot represents one biological replicate. ** p < 0.01; **** p < 0.0001.
Fig 5: Effect of IGF-1 on the proliferative and invasive properties of LNCaP cells. LNCaP cells were treated with IGF-1 (150 ng/mL) for 72 h or left untreated (control). Cell proliferation was assessed by (a) cell counting and (b) colony formation assay. Invasive potential was evaluated by (c) an invasion assay and by analyzing the transcript levels of the invasion-related matrix metalloproteinases (d) MMP-1, MMP-7, and MMP-9. Data are presented as the mean ± standard deviation (SD) of three independent experiments; each black dot represents one biological replicate * p < 0.05; *** p < 0.001; **** p < 0.0001 versus untreated cells.
Supplier Page from Thermo Fisher Scientific for Human IGF-1 ELISA Kit