Journal: International Journal of Molecular Sciences

Article Title: Estrogen α and β Receptor Expression in the Various Regions of Resected Glioblastoma Multiforme Tumors and in an In Vitro Model

doi: 10.3390/ijms25074130

Figure Lengend Snippet: Expression of ERα gene ( A ) and ERα protein ( B ) in U87 cells cultured under different conditions. Data are representative of each group cultured in control, nutrient-deficient, hypoxic, and necrotic conditions. Statistical analysis was performed using t -test *, p < 0.005.

Article Snippet: Cell culture of the U87 line (obtained from the European Collection of Authenticated Cell Cultures (ECACC)) was performed under standard conditions of 37 °C, 95% humidity, and 5% CO 2 , according to the manufacturer’s instructions.

Techniques: Expressing, Cell Culture, Control

Journal: International Journal of Molecular Sciences

Article Title: Estrogen α and β Receptor Expression in the Various Regions of Resected Glioblastoma Multiforme Tumors and in an In Vitro Model

doi: 10.3390/ijms25074130

Figure Lengend Snippet: Representative images taken with the FV1000 confocal microscope system (Olympus, Hamburg, Germany) show ERα protein expression in U87 cells cultured under specific conditions: control ( A ), nutrient deficiency ( B ), hypoxia ( C ), and necrotic conditions ( D ). FITC (AR) and DAPI (nuclear) markers were used. Microphotographs were taken at ×20 magnification ( A , B , D ) and ×40 magnification ( C ); scale bar 30 µm.

Article Snippet: Cell culture of the U87 line (obtained from the European Collection of Authenticated Cell Cultures (ECACC)) was performed under standard conditions of 37 °C, 95% humidity, and 5% CO 2 , according to the manufacturer’s instructions.

Techniques: Microscopy, Expressing, Cell Culture, Control

Journal: International Journal of Molecular Sciences

Article Title: Estrogen α and β Receptor Expression in the Various Regions of Resected Glioblastoma Multiforme Tumors and in an In Vitro Model

doi: 10.3390/ijms25074130

Figure Lengend Snippet: Expression of the ERβ gene ( A ) and ERβ protein ( B ) in U87 cells cultured under different conditions. Data are representative of each group cultured in control, nutrient-deficient, hypoxic, and necrotic conditions. Statistical analysis was performed using a t -test * p < 0.05.

Article Snippet: Cell culture of the U87 line (obtained from the European Collection of Authenticated Cell Cultures (ECACC)) was performed under standard conditions of 37 °C, 95% humidity, and 5% CO 2 , according to the manufacturer’s instructions.

Techniques: Expressing, Cell Culture, Control

Journal: International Journal of Molecular Sciences

Article Title: Estrogen α and β Receptor Expression in the Various Regions of Resected Glioblastoma Multiforme Tumors and in an In Vitro Model

doi: 10.3390/ijms25074130

Figure Lengend Snippet: Representative images taken with the FV1000 confocal microscope system (Olympus, Hamburg, Germany) show ERβ protein expression in U87 cells cultured under specific conditions: control ( A ), nutrient deficiency ( B ), hypoxia ( C ), and necrotic conditions ( D ). FITC (AR) and DAPI (nuclear) markers were used. Microphotographs were taken at ×20 magnification ( A , B , D ) and ×40 magnification ( C ); scale bar 30 µm.

Article Snippet: Cell culture of the U87 line (obtained from the European Collection of Authenticated Cell Cultures (ECACC)) was performed under standard conditions of 37 °C, 95% humidity, and 5% CO 2 , according to the manufacturer’s instructions.

Techniques: Microscopy, Expressing, Cell Culture, Control

Journal: Translational Oncology

Article Title: The role of A-kinase interacting protein 1 in regulating progression and stemness as well as indicating the prognosis in glioblastoma

doi: 10.1016/j.tranon.2022.101463

Figure Lengend Snippet: AKIP1 mRNA and protein expression in GBM and control cell lines. AKIP1 mRNA expression (A) and protein expression (B,C) in GBM cell lines (including GB-1, U-251 MG, LN18, A172, U-87 MG) and control cell lines. Each experiment was replicated three times.

Article Snippet: Human GBM cell lines, including GB1, U-251 MG, LN18, A172, and U-87 MG, were purchased from the American Type Culture Collection (ATCC, USA) or Japanese Collection of Research Bioresources Cell Bank (JCRB, Japan), respectively.

Techniques: Expressing, Control

Journal: Oncotarget

Article Title: Type 5 phosphodiesterase regulates glioblastoma multiforme aggressiveness and clinical outcome

doi: 10.18632/oncotarget.14656

Figure Lengend Snippet: A . Wound healing assay and B . Histogram showing the mean percentage ±SD of closure of sh-scrT98G or sh1 or sh2-PDE5 cells monolayers in the presence or absence of 1 μM sildenafil or 100 μM 8br-cGMP. C . Migration assay on matrigel cushions through transwell filters of sh-scr or sh1-PDE5 T98G cells in the presence or absence of 1 μM sildenafil or 100 μM 8br-cGMP. Scale bars= 50 μm. D . Histogram showing the relative migration capacity (fold increase ± SD) of cells that passed through the filter in the different treatments compared to control cells. E . Migration assay on matrigel cushions through transwell filters of mock- or PDE5-overexpressing U87MG (upper panels) or GBM line 83 cells (lower panels). Scale bar= 20 μm. F . Histogram showing the mean percentage ± SD of cells that passed through the filter. G . Histogram of the mean percentage ±SD of secreted MMP-2 in gelatin zymography (bottom image) in sh-scr or sh1-PDE5 T98G and H . in mock- or PDE5-overexpressing U87MG cells and I . in sh-scr T98G stimulated with CNP, GSNO or sildenafil at the indicated concentrations. Data have been obtained from three independent experiments. (*p < 0.05; **p < 0.001).

Article Snippet: The GBM cell lines (T98G, U87MG, A172, U251, LN18) used in this study were grown in DMEM, (Euroclone; Milan, Italy) with 10% fetal bovine serum (FBS) (Euroclone).

Techniques: Wound Healing Assay, Migration, Control, Zymography

Journal: Oncotarget

Article Title: Type 5 phosphodiesterase regulates glioblastoma multiforme aggressiveness and clinical outcome

doi: 10.18632/oncotarget.14656

Figure Lengend Snippet: A . PKG1 low and B . PKG1 high GBM samples, as revealed by immuno-histochemistry. Black arrows point to PKG1 expression within the vessel walls. White arrow points to tumor cells. Scale bar=20μm. C . Expression levels of PKGII, PKG1α and PKG1β and total PKG1 in sh-scr and sh1-PDE5 T98G cells and in wtT98G, U87MG, A172, LN18, 83 GBM lines. D . pVASP levels in sh-scr or sh1-PDE5 T98G cells before and after Y-27632, 8br-cGMP (50 μM) treatments. Representative image out of 4 independent experiments. E . Phosphorylation levels of MYPT1 (pSer695) in sh-PDE5 or sh1-scr T98G upon the indicated treatments [8br-cGMP, 50 μM; Y-27632 (RI), 10 μM); KT5823 (PKGi), 2 μM]. F . Densitometric analysis of western blots from three independent experiments is shown. Bars represent the mean ±SD. G . sh-scr and sh1-PDE5 T98G invasion assays on matrigel cushions in the presence of Y-27632 or 8br-cGMP or both. H . Histogram showing the relative migration capacity (fold increase ± SD) of cells that reached the bottom of the matrigel cushion in the different treatments compared to control cells. (**p < 0.001).

Article Snippet: The GBM cell lines (T98G, U87MG, A172, U251, LN18) used in this study were grown in DMEM, (Euroclone; Milan, Italy) with 10% fetal bovine serum (FBS) (Euroclone).

Techniques: Immunohistochemistry, Expressing, Phospho-proteomics, Western Blot, Migration, Control

Journal: Oncotarget

Article Title: Type 5 phosphodiesterase regulates glioblastoma multiforme aggressiveness and clinical outcome

doi: 10.18632/oncotarget.14656

Figure Lengend Snippet: A . sh-scr (PDE5+) or sh1-PDE5 (PDE5-) T98G cells were treated with increasing X-ray doses from 2 to 4 Gy and probed with anti-PAR and anti-PARP-1 antibodies. B . sh-scr or sh1-PDE5 T98G cells were exposed to X-rays at 4 Gy in the presence or absence (+/-) of the PARP1 inhibitor olaparib (1 μM). Total protein PARylation levels were assayed after 24 h. C . Schematic representation of cGMP signaling pathway promoting migration and DNA repair in GBM cells.

Article Snippet: The GBM cell lines (T98G, U87MG, A172, U251, LN18) used in this study were grown in DMEM, (Euroclone; Milan, Italy) with 10% fetal bovine serum (FBS) (Euroclone).

Techniques: Migration

Journal: Heliyon

Article Title: Dishevelled-associated antagonist of β-catenin homolog 3 (DACT3) suppresses glioma progression though Notch1 signaling pathway in β-catenin-dependent manner

doi: 10.1016/j.heliyon.2023.e23511

Figure Lengend Snippet: Overexpression of DACT3 suppresses progression of glioma cells in vitro.A. Western blot analysis shows endogenous expression of DACT3 in four glioma cell lines (U87, U118, T98, U251, n=3, The full, non-adjusted images are shown in Supplement file). B. Expression levels of DACT3 proteins in four glioma cell lines were quantified and normalized to GAPDH levels (n=3). C. Western blot analysis demonstrates DACT3 over-expression in U118 and U251 glioma cells (n=3, The full, non-adjusted images are shown in Supplement file). D. Expression levels of DACT3 proteins in DACT3 over-expressing glioma cells were quantified and normalized to GAPDH levels (n=3). E-J. Cell proliferation was detected in U251 and U118 glioma cells by CCK-8 assay ( E. F ), colony formation assay ( G. H ) and Ki67 immunofluorescence assay ( I. J ) (400×, n=3). K-N. Invasive ( K. L ) and migrative ( M. N ) ability are tested in U251 and U118 glioma cells by transwell assay (400×, n=3). O. P. Adhesion ability is tested in U251 and U118 glioma cells by Cell adhesion assay (400×, n=3). DACT3: Dishevelled-associated antagonist of β-catenin homolog 3; ctrl: transfected with empty vectors; overDACT3: transfected with DACT3vectors. (* P <0.05, ** P < 0.01, *** P < 0.001; * ctrl vs overDACT3).

Article Snippet: GBM cell line T98 (CL-0583) was obtained from Procell Life Science &Technology (Wuhan, China).

Techniques: Over Expression, In Vitro, Western Blot, Expressing, CCK-8 Assay, Colony Assay, Immunofluorescence, Transwell Assay, Cell Adhesion Assay, Transfection