Journal: Frontiers in Oncology

Article Title: The role of tumor-associated macrophages in glioma cohort: through both traditional RNA sequencing and single cell RNA sequencing

doi: 10.3389/fonc.2023.1249448

Figure Lengend Snippet: (A) The cck8 assay in U87 cells; (B) The cck8 assay in U251 cells; (C) The Edu assay in U87 cells; (D) The Edu assay in U251 cells.

Article Snippet: Human glioma cell lines U251 and U87 were purchased from Beijing Zhongyuan Heju Biotechnology Co., Ltd. (Beijing, China).

Techniques: CCK-8 Assay, EdU Assay

Journal: Frontiers in Molecular Neuroscience

Article Title: Subcellular Localization of Sprouty2 in Human Glioma Cells

doi: 10.3389/fnmol.2019.00073

Figure Lengend Snippet: Immunofluorescence against Sprouty2 in glioma cell lines. As compared to naive U251 cells (a) stronger labeling is observed in U87 (d) and SF126 cells (g) Immunostaining of Sprouty2 overexpressing cells reveals prominent labeling of peripheral regions, particularly in cellular protrusions and ruffles ( c,f,i ; arrows). Reduced but not absent labeling is observed in cells with shRNA-mediated Sprouty2 down-regulation (b,e,h) which was confirmed by quantitative Western blotting experiments (j) demonstrating specific protein bands in naive, shRNA expressing or Sprouty2 overexpressing cells (letters correspond to cells shown in respective images above).

Article Snippet: The standard glioma cell lines U251, U87 and SF126 were authenticated by short tandem repeat analysis (Microsynth) and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma) or in Roswell Park Memorial Institute (RPMI) medium supplemented with 10% Fetal Bovine Serum (FBS, Gibco), 1% antibiotic-antimycotic (Gibco) and 2 mM L-glutamine (Gibco).

Techniques: Immunofluorescence, Labeling, Immunostaining, shRNA, Western Blot, Expressing

Journal: Frontiers in Molecular Neuroscience

Article Title: Subcellular Localization of Sprouty2 in Human Glioma Cells

doi: 10.3389/fnmol.2019.00073

Figure Lengend Snippet: Immunogoldelectron microscopy of U251, SF126 and U87 glioma cells transiently expressing NG-tagged Sprouty2. In U251 (a) and SF126 cells (b) anti-NG-Sprouty2 label (arrows) is found both at the limiting membrane and inside of late endosomes (in addition to its cytoplasmic distribution), whereas the label in U87 cells ( c , arrowheads) appears predominantly in the cytoplasm. Anti-Sprouty2 antibodies detect cytoplasmic Sprouty2 in U251 cells (d) and at the limiting membrane of late endosomes in SF126 cells (e) . LE = late endosome; M = mitochondrion; N = nucleus.

Article Snippet: The standard glioma cell lines U251, U87 and SF126 were authenticated by short tandem repeat analysis (Microsynth) and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma) or in Roswell Park Memorial Institute (RPMI) medium supplemented with 10% Fetal Bovine Serum (FBS, Gibco), 1% antibiotic-antimycotic (Gibco) and 2 mM L-glutamine (Gibco).

Techniques: Microscopy, Expressing, Membrane

Journal: Frontiers in Molecular Neuroscience

Article Title: Subcellular Localization of Sprouty2 in Human Glioma Cells

doi: 10.3389/fnmol.2019.00073

Figure Lengend Snippet: Distinct association of endogenous Sprouty2 with vimentin (intermediate) filaments in U251 (a–c) , SF126 (d–f) and U87 cells (g–i) . Note the punctate staining of Sprouty2 and vimentin in a subpopulation of SF126 cells (treated with EGF-647). However, other SF126 cells (see ), as well as U251 and U87 cells, exhibited the expected filamentous vimentin staining with associated Sprouty2 immunofluorescence (yellow structures in merged images).

Article Snippet: The standard glioma cell lines U251, U87 and SF126 were authenticated by short tandem repeat analysis (Microsynth) and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma) or in Roswell Park Memorial Institute (RPMI) medium supplemented with 10% Fetal Bovine Serum (FBS, Gibco), 1% antibiotic-antimycotic (Gibco) and 2 mM L-glutamine (Gibco).

Techniques: Staining, Immunofluorescence

Journal: Frontiers in Molecular Neuroscience

Article Title: Subcellular Localization of Sprouty2 in Human Glioma Cells

doi: 10.3389/fnmol.2019.00073

Figure Lengend Snippet: Confocal microscopy of endogenous Sprouty2 (a) and vimentin (a’) followed by super-resolution (STED) microscopy (b–d) reveals endosomal ( b , inset with higher magnification of spots) and punctate labeling ( b , arrow) in vimentin rich perinuclear areas of SF126 cells ( b’ ). Sprouty2 spots are <50 nm in diameter and partially associated with vimentin filaments (c) . Anti-FLAG immunofluorescence against C-terminally truncated FLAG-Sprouty2 demonstrates that Sprouty2 spots (presumably oligomers) form independently of the C-terminus (d) . Immunoprecipitation of Sprouty2-FLAG in U251 cells confirms the association of Sprouty2 with vimentin but not with tubulin (e) . Anti-FLAG immunoprecipitates (IP) were analyzed in comparison with total cell lysates (TCL) by immunoblotting (IB) probed with antibodies against vimentin, tubulin and FLAG.

Article Snippet: The standard glioma cell lines U251, U87 and SF126 were authenticated by short tandem repeat analysis (Microsynth) and cultured in Dulbecco’s Modified Eagle’s Medium (DMEM, Sigma) or in Roswell Park Memorial Institute (RPMI) medium supplemented with 10% Fetal Bovine Serum (FBS, Gibco), 1% antibiotic-antimycotic (Gibco) and 2 mM L-glutamine (Gibco).

Techniques: Confocal Microscopy, Microscopy, Labeling, Immunofluorescence, Immunoprecipitation, Comparison, Western Blot

Journal: Nutrients

Article Title: Inhibitory Effects of Urolithins, Bioactive Gut Metabolites from Natural Polyphenols, against Glioblastoma Progression

doi: 10.3390/nu15234854

Figure Lengend Snippet: Aryl hydrocarbon receptor (AhR) expression in GBM is correlated with pathologic grades of human glioma and involves adhesion molecule expression. ( A ) AhR mRNA levels of patients’ specimens from human glioma microarray data set GSE4290. ( B ) U251 human GBM was treated with AhR antagonist 6,2′,4′-trimethoxyflavone (TMF) and stimulated with tumor necrosis factor (TNF)-α for another 24 h. Vascular cell adhesion molecule (VCAM)-1 expression was evaluated by Western blotting. ( C ) U251 and U87 GBM cells were treated with TMF for 30 min and stimulated with TNF-α for another 24 h. The cultured medium was replaced with fresh serum-free medium. Supernatants were collected after 24 h. Soluble VCAM-1 expression was determined using enzyme-linked immunosorbent assay. ( D ) U251 GBM cells were pretreated with TMF for 30 min and stimulated with TNF-α for another 24 h. BCECF-AM-labeled THP-1 cells were added to GBM cells and cultured for 30 min. THP-1 adhesion to GBM cells was observed under a fluorescence microscope. Quantitative results are shown in ( E ). ( F ) U251 human GBM cells were treated with urolithin A (10 μM), TMF (10 μM), or cotreatment for 24, 48, and 72 h. Cell viability was performed using MTT assay. ( G , H ) U251 human GBM cells were treated with urolithins and TMF for 24 h; cell migration was detected by using transwell assay. Each bar represents the mean ± standard error of the mean. * p < 0.05 compared with the control group. # p < 0.05 compared with the TNF-α group.

Article Snippet: U87 human glioma, ALTS1C1 mouse glioma, and THP-1 human monocytes were obtained from the Bioresource Collection and Research Center (BCRC No. 60360, 60430 and 60582; Hsinchu, Taiwan).

Techniques: Expressing, Microarray, Western Blot, Cell Culture, Enzyme-linked Immunosorbent Assay, Labeling, Fluorescence, Microscopy, MTT Assay, Migration, Transwell Assay, Control

Journal: Nutrients

Article Title: Inhibitory Effects of Urolithins, Bioactive Gut Metabolites from Natural Polyphenols, against Glioblastoma Progression

doi: 10.3390/nu15234854

Figure Lengend Snippet: Inhibition of aryl hydrocarbon receptor using 6,2′,4′-trimethoxyflavone (TMF) represses human macrophage condition medium (HMCM)-induced programmed death ligand (PD-L1) expression in GBM cells. U251 ( A ) and U87 ( B ) human GBM were treated with TMF for 30 min and stimulated with HMCM for another 24 h. PD-L1 expression was determined using Western blotting. Quantitative results are shown in ( C ). Each bar represents the mean ± standard error of the mean ( n = 3). * p < 0.05 compared with the control group. # p < 0.05 compared with the HMCM group.

Article Snippet: U87 human glioma, ALTS1C1 mouse glioma, and THP-1 human monocytes were obtained from the Bioresource Collection and Research Center (BCRC No. 60360, 60430 and 60582; Hsinchu, Taiwan).

Techniques: Inhibition, Expressing, Western Blot, Control