Journal: Molecules

Article Title: Antitumoral Activity of the Universal Methyl Donor S -Adenosylmethionine in Glioblastoma Cells

doi: 10.3390/molecules29081708

Figure Lengend Snippet: Effect of AdoMet on cell viability of GBM cells. U251MG, U87MG, U343MG, and NHA cell lines were treated or not (control) with increasing amounts of AdoMet (72–1000 μM) (X axis in log scale) for 24, 48, and 72 h. Cell viability was then measured by MTT assay and expressed as a percentage of the control cells. Error bars depict the standard deviation (SD) of triplicated measurements and are representative of three separate experiments, ** p < 0.01, **** p < 0.001 versus untreated cells.

Article Snippet: U343MG stabilized cell line was purchased from CLS Cell Lines Service GmbH (Eppelheim, Germany, Catalog No. 300365).

Techniques: Control, MTT Assay, Standard Deviation

Journal: Molecules

Article Title: Antitumoral Activity of the Universal Methyl Donor S -Adenosylmethionine in Glioblastoma Cells

doi: 10.3390/molecules29081708

Figure Lengend Snippet: Effects of AdoMet on cell cycle distribution in GBM cells. ( A ) U251MG, U87MG, and U343MG cells were cultured for 72 h in medium supplemented or not (control) with AdoMet 500 μM, and the cell cycle was then assessed by flow cytometry. Histograms show the percentage of cells in each phase of cell cycle. At least 2 × 10 4 events were analyzed for each sample. Data represent the average of three independent experiments. Error bars depict the SDs. * p < 0.05 versus untreated cells. ( B ) The levels of the main cell cycle regulatory proteins were evaluated by Western blot, and the relative densitometric analyses performed for each protein in relation to its relative housekeeping protein were reported as a percentage of untreated cells (100%). Uncropped images of Western blots and their housekeeping proteins are reported in . Representative housekeeping β-actin protein used as a loading control was shown. Error bars represent the SD. * p < 0.05, ** p < 0.01, *** p < 0.005 versus untreated cells. The images are representative of three immunoblotting analyses obtained from three independent experiments.

Article Snippet: U343MG stabilized cell line was purchased from CLS Cell Lines Service GmbH (Eppelheim, Germany, Catalog No. 300365).

Techniques: Cell Culture, Control, Flow Cytometry, Western Blot

Journal: Molecules

Article Title: Antitumoral Activity of the Universal Methyl Donor S -Adenosylmethionine in Glioblastoma Cells

doi: 10.3390/molecules29081708

Figure Lengend Snippet: Effects of AdoMet on apoptosis in GBM cells. ( A ) U251MG, U87MG, and U343MG cells were cultured for 72 h in medium supplemented or not (control) with AdoMet 500 μM, and the apoptotic process was then assessed by FACS analysis. Histograms show the percentage of apoptotic cells. For each sample, at least 2 × 10 4 events were analyzed. Data represent the average of three independent experiments. Error bars depict the SDs. * p < 0.05 versus untreated cells. ( B ) The protein levels of uncleaved caspase-3 and PARP-1 and the corresponding cleaved forms were evaluated by Western blot, and the relative densitometric analyses are reported as percentage of untreated control (100%). Housekeeping proteins, used as a loading control, were reported. Error bars represent the SDs. ** p < 0.01 ***, p < 0.005, **** p < 0.001 versus untreated cells. The images are representative of three immunoblotting analyses obtained from three independent experiments. Uncropped images of Western blots and their housekeeping proteins are reported in .

Article Snippet: U343MG stabilized cell line was purchased from CLS Cell Lines Service GmbH (Eppelheim, Germany, Catalog No. 300365).

Techniques: Cell Culture, Control, Western Blot

Journal: Molecules

Article Title: Antitumoral Activity of the Universal Methyl Donor S -Adenosylmethionine in Glioblastoma Cells

doi: 10.3390/molecules29081708

Figure Lengend Snippet: Effects of AdoMet on DNA damage response in GBM cells. ( A ) U251MG, U87MG, and U343MG cells were cultured for 72 h in medium supplemented or not (control) with AdoMet 500 μM. The levels of the main proteins involved in DNA repair and DNA damage response were then evaluated by Western blot, and the relative densitometric analyses, performed for each protein in relation to its relative housekeeping included in the (showing the uncropped images of Western blots), are reported as a percentage of untreated cells (100%). Representative housekeeping β-actin protein, used as a loading control, was reported. Error bars represent the SD. * p < 0.05, ** p < 0.01, *** p < 0.005 versus untreated cells. ( B ) Effect of AdoMet on γH2AX and RAD51 foci formation. Representative images of immunofluorescence staining for phosphorylated γH2AX (red) and RAD51 (green) in U343MG cells treated or not (control) with AdoMet. The overlapping of the two signals (merge panel) in AdoMet-treated cells evidenced the reduced expression of RAD51 and the concomitant increase in γH2AX foci. Hoechst 33258 was used for nuclear staining (blue). Scale bar: 20 μm. About 100 nuclei for each group were scored in each experiment, and a threshold of five foci per cell was considered positive and reported in the histograms as a mean of γH2AX and RAD51 foci. Values represent the means of three experiments ± SD (* p < 0.05, ** p < 0.01).

Article Snippet: U343MG stabilized cell line was purchased from CLS Cell Lines Service GmbH (Eppelheim, Germany, Catalog No. 300365).

Techniques: Cell Culture, Control, Western Blot, Immunofluorescence, Staining, Expressing