Journal: Biology

Article Title: Overexpression of Lipocalin-2 Inhibits Proliferation and Invasiveness of Human Glioblastoma Multiforme Cells by Activating ERK Targeting Cathepsin D Expression

doi: 10.3390/biology10050390

Figure Lengend Snippet: CTSD expression is inhibited by LCN2 overexpression and correlates inversely with GBM patient survival. ( A ) A human protease array profile was performed on whole cell lysate from LCN2-overexpressed or Neo-GBM8901 and U-251 cells. ( B ) The expression of protein and ( C ) mRNA of LCN2-overexpressed or Neo-GBM8901 and U-251 cells were analyzed through western blot and RT-qPCR, respectively. ( D , G ) Comparison of CTSD expression level between GBM and normal tissues of GBM patients from the GEPIA and GSE4290 database. β-actin as protein loading internal control. GAPDH as mRNA control. Comparison of the overall survival ( E , H ) and disease-free survival ( F ) between low and high expression of CTSD from the GEPIA and GSE13041 database. Data are presented as the mean ± SE of at least three independent experiments. * p < 0.05, ** p < 0.01 compared with the Neo cells or normal tissues.

Article Snippet: U0126 (MEK inhibitor), siRNA-CTSD (si-CTSD), Cathepsin D (CTSD), t-ERK, β-actin, HRP-mouse and HRP-rabbit antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Expressing, Over Expression, Western Blot, Quantitative RT-PCR, Comparison, Control

Journal: Biology

Article Title: Overexpression of Lipocalin-2 Inhibits Proliferation and Invasiveness of Human Glioblastoma Multiforme Cells by Activating ERK Targeting Cathepsin D Expression

doi: 10.3390/biology10050390

Figure Lengend Snippet: Effect of LCN2 overexpression on ERK-targeted CTSD expression. ( A ) Western blot analysis of total lysates of LCN2-overexpressing or Neo-GBM8901 and U-251 cells to determine the protein expression of LCN2, p-ERK, and t-ERK (left panel). ( B ) Western blot analysis of the same cells were treated with the MEK inhibitor U0126 (20 μM) for 2 h to determine the protein expression of LCN2, p-ERK, t-ERK, and CTSD (right panel). ( C ) The migration and invasiveness of cells following treatment with or without U0126 (20 µM) in LCN2-overexpressing or Neo-U-251 cells. ( D ) Transfection with si-Con or si-CTSD in LCN2-overexpressing or Neo-U-251 cells for 48 h, then extracted the total proteins and measured by western blot. ( E ) The migration and invasiveness of cells following transfected with si-Con or si-CTSD in LCN2-overexpressing or Neo-U-251 cells. β-actin was used as an internal control for protein loading Data are presented as the mean ± SE of at least three independent experiments. * p < 0.05, ** p < 0.01 compared with Neo cells or si-Con in Neo overexpressing cells. # p < 0.05, compared with LCN2-overexpressing cells or si-Con in LCN2 overexpressing cells.

Article Snippet: U0126 (MEK inhibitor), siRNA-CTSD (si-CTSD), Cathepsin D (CTSD), t-ERK, β-actin, HRP-mouse and HRP-rabbit antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Over Expression, Expressing, Western Blot, Migration, Transfection, Control