Journal: Radiation Oncology (London, England)

Article Title: Integrative analysis of therapy resistance and transcriptomic profiling data in glioblastoma cells identifies sensitization vulnerabilities for combined modality radiochemotherapy

doi: 10.1186/s13014-022-02052-z

Figure Lengend Snippet: mRNA expression analysis of DNA damage response (DDR) regulator genes, and MGMT promoter methylation analysis in human GBM cell lines. A Basal mRNA expression levels of DDR regulator genes in GBM cell lines A172, LN18, LN229, T98G, U87-MG, U138-MG, and U251-MG as measured by qRT-PCR (ddCT method). Expression levels were normalized to a matrix of three reference genes (18S rRNA, β2-Microglobulin, and 5’-Aminolevulinate Synthase-1), and calibrated on the expression levels of astrocytes. Three replicates were analyzed per cell line. Expression values (log2-transformed) and samples were subjected to unsupervised hierarchical clustering. B Correlation of MGMT promoter methylation status and MGMT mRNA expression levels in the employed GBM cell lines as detected by methylome array and qRT-PCR, respectively. C Protein expression levels of MGMT in GBM cells as measured by western blot. Vinculin served as loading control

Article Snippet: The human GBM cell lines A172, LN18, LN229, and T98G were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA), and the human GBM cell lines U87-MG, U138-MG, and U251-MG were purchased from Cell Lines Service GmbH (CLS, Eppelheim, Germany).

Techniques: Expressing, Methylation, Quantitative RT-PCR, Transformation Assay, Western Blot

Journal: Radiation Oncology (London, England)

Article Title: Integrative analysis of therapy resistance and transcriptomic profiling data in glioblastoma cells identifies sensitization vulnerabilities for combined modality radiochemotherapy

doi: 10.1186/s13014-022-02052-z

Figure Lengend Snippet: Validation of candidate genes by targeted inhibition and clonogenic survival assays. A Clonogenic survival of A172 and U251-MG cells after single-shot IR (0–10 Gy) and ATR inhibition by AZD-6738 (0–1.0 μM, upper panel). Immunofluorescence staining of γH2AX and 53BP1 20 h after irradiation at 4 Gy ± ATR inhibition by 1 µM AZD-6738 (lower panel). γH2AX is depicted in green, 53BP1 in red, and DNA is depicted in blue. Scale bar represents 10 µm. B Clonogenic survival of A172 and U251-MG cells after single-shot IR and LIG4 inhibition by L189 (0–50 μM, upper panel). Immunofluorescence staining of γH2AX and 53BP1 20 h after irradiation at 4 Gy ± LIG4 inhibition by 50 µM L189 as performed in A (lower panel). C Clonogenic survival of A172, U138-MG, and U251-MG cells after 24 h treatment with TMZ (0–200 μM) and ATM inhibition by KU-60019 (0–1.0 μM). Super-imposed fitting functions are linear-quadratic in case of radiation experiments and logistic in case of TMZ treatment. Results of three independent experiments are depicted for each condition, and p-values were calculated by two-way ANOVAs

Article Snippet: The human GBM cell lines A172, LN18, LN229, and T98G were purchased from American Type Culture Collection (ATCC, Manassas, VA, USA), and the human GBM cell lines U87-MG, U138-MG, and U251-MG were purchased from Cell Lines Service GmbH (CLS, Eppelheim, Germany).

Techniques: Inhibition, Immunofluorescence, Staining, Irradiation

Journal: Trends in cell biology

Article Title: Mitochondrial oxidative phosphorylation TRAP(1)ped in tumor cells

doi: 10.1016/j.tcb.2014.03.005

Figure Lengend Snippet: TRAP1 expression levels in different tissue or cell types, either normal (upper panel) or transformed (lower panel)

Article Snippet: U-138 MG, U-251 MG, U-87 MG , Medium (IHC) , The Human Protein Atlas.

Techniques: Expressing, Transformation Assay, Real-time Polymerase Chain Reaction

Journal: Biomolecules

Article Title: ADAR2 Protein Is Associated with Overall Survival in GBM Patients and Its Decrease Triggers the Anchorage-Independent Cell Growth Signature

doi: 10.3390/biom12081142

Figure Lengend Snippet: ADAR2 correlates with patients’ overall survival. ( a ) IHC analysis (H&E and the corresponding ADAR2 antibody staining) of two glioblastoma tissues (examples of high and low ADAR2 levels). Subpanels A and B show a representative GBM case with high ADAR2 expression; in subpanels C and D, a GBM case with low ADAR2 expression is shown; scale bar: 200 µm. ( b ) Kaplan–Meier curve comparing the survival of GBM patients (n = 39) stratified by ADAR2 levels. The red and blue lines represent low and high ADAR2 expression, respectively, following the scores indicated in Material and Methods ( p = 0.0005; HR 3.954; 95% CI from 1834 to 8525).

Article Snippet: We analyzed the endogenous ADAR2 level (mRNA and protein) in several human GBM cell lines (A172, T98G, U118MG, U87MG, LN18, and U138-MG) that show diverse abilities to give rise to a growing tumor mass if implanted in a mouse model ( www.atcc.org , accessed on 1 January 2022) ( a,b).

Techniques: Staining, Expressing

Journal: Biomolecules

Article Title: ADAR2 Protein Is Associated with Overall Survival in GBM Patients and Its Decrease Triggers the Anchorage-Independent Cell Growth Signature

doi: 10.3390/biom12081142

Figure Lengend Snippet: ADAR2 expression level and activity across GBM cell lines. ( a ) ADAR2 expression (qRT-PCR) in normal brain (fetal and adult), astrocytes, and glioblastoma cell lines (T98G, U138-MG, U87-MG, U118-MG, LN-18, and A172). Ct values were normalized to GAPDH mRNA levels. Mean ± standard deviation ( n = 3), values are representative as means ± SD, * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001. Of note, ADAR2 activity is more robust in the whole brain (either adult or fetal) because of neuronal cells, where ADAR2 is highly active. ( b ) Western blotting analysis showing the ADAR2 level in glioblastoma cell lines. No control was added as the ADAR2 level was too high. ( c ) The recoding editing index (REI) in normal astrocytes (×10), T98-G, U87-MG, U118-MG, LN-18, and A172 glioblastoma cells lines is shown. REI values were calculated as the weighted average of editing levels over all known recording sites from the highly accurate list.

Article Snippet: We analyzed the endogenous ADAR2 level (mRNA and protein) in several human GBM cell lines (A172, T98G, U118MG, U87MG, LN18, and U138-MG) that show diverse abilities to give rise to a growing tumor mass if implanted in a mouse model ( www.atcc.org , accessed on 1 January 2022) ( a,b).

Techniques: Expressing, Activity Assay, Quantitative RT-PCR, Standard Deviation, Western Blot, Control