Journal: Journal of Cell Science

Article Title: Interplay between nuclear survivin and the PRC2 complex and its impact on H3K27me3-directed transcriptional repression

doi: 10.1242/jcs.264572

Figure Lengend Snippet: Hypoxia increases expression of EZH2, H3K27me3 and survivin. (A) Immunoblots of WCEs from U2OS, HeLa and MRC5 lines cultured under normoxic or hypoxic environments (24 h). Blots were immunoprobed with anti-EZH2, anti-H3K27me3 and anti-survivin antibodies. Anti-Hif1a used to prove the hypoxic state had been induced, and anti-tubulin was used as a loading control. (B–D) Quantification of immunoblots represented in A from three independent experiments demonstrating that EZH2, H3K27me3 and survivin are all more abundant under hypoxia. Data presented are means±s.d. * P <0.05, ** P <0.01, *** P <0.001 (two-way ANOVA with Tukey's multiple comparisons post test).

Article Snippet: Human cervical cancer cells (HeLa), human bone osteosarcoma cells (U2OS), retinal pigment epithelial cells (RPE) and human breast cancer cells (MCF7) were originally from ATCC.

Techniques: Expressing, Western Blot, Cell Culture, Control

Journal: Nature Communications

Article Title: Over four minutes of pyruvate T 1 using chemically and physically induced deceleration of relaxation

doi: 10.1038/s41467-026-73214-w

Figure Lengend Snippet: Three dissolution media compositions were tested: D 2 O filtr. being like sample #C, D 2 O like #C without the filtration step, and H 2 O being like sample #G. A The retained polarization of pyruvate after a 45 s transfer measured at 1 T without cells was (29.9 ± 1.7)%, (35.6 ± 1.7)%, and (25.8 ± 1.8)% for the samples as introduced. B The polarization lifetime of pyruvate, corrected for flip-angle and measured at 1 T without cells, was (162.1 ± 2.8) s, (153.8 ± 10.1) s, and (77.1 ± 1.2) s for the samples as introduced, respectively. C The observed maximum SNR for the metabolic product lactate in HeLa cells was (227.9 ± 9.4), (365.3 ± 27.4), and (156.7 ± 14.6), respectively, using the same acquisition parameters. A transfer magnet of about 35 mT was used for D 2 O filtr. and D 2 O, but not for H 2 O. The transfer time was kept constant across all samples to accommodate about 40 s for the transfer, including filtration. D 2 O and H 2 O experiments were done on the same day. pH of the dissolution medium (and cells) was 7.95 ± 0.16 (7.65 ± 0.24), 7.38 ± 0.08 (7.10 ± 0.03), and 7.65 ± 0.03 (7.5 ± 0.01), respectively. Two biological repeats were performed for D 2 O filtr., and three for D 2 O and H 2 O, repetition was performed by growing independent flasks for each experiment. H 2 O is treated as the control group. Respective kinetics in Supplementary Note , Supplementary Fig. . The bar plots show mean values ± sample SD. Source data are provided as a Source Data file.

Article Snippet: The human cervical cancer HeLa cells (from ATCC: CCL-2) were maintained at 37 °C with 5% CO2 in high glucose Dulbecco’s Modified Eagle Medium (DMEM; Gibco, Thermo Fisher Scientific, Waltham, MA, USA) containing 4.5 g/L D -glucose, L -glutamine, 1 mM sodium pyruvate (Gibco), 10% fetal bovine serum (FBS South America; Pan-Biotech, Aidenbach, Germany), and 1% penicillin/streptomycin (Gibco).

Techniques: Dissolution, Filtration, Control

Journal: International Journal of Molecular Sciences

Article Title: Reactivation of P53 Antiproliferative and Pro-Apoptotic Pathways by Resveratrol in Mutant P53 Cancer Cell Lines

doi: 10.3390/ijms27104481

Figure Lengend Snippet: Dose-dependent effect of resveratrol (RES) on cellular metabolic activity in cancer cell lines with different p53 status. Cells were treated with increasing concentrations of RES (50–250 µM) for 48 h. Cellular metabolic activity was assessed by means of MTT assay and normalized to that of vehicle-treated controls (EtOH 0.3% v / v ). Data represent mean ± SD of at least four independent experiments. IC50 values are indicated for each cell line (presented in box).

Article Snippet: The human cervical cancer cell line C33A ( p53 mutation R273C) was purchased from the American Type Culture Collection (ATCC cat. no. HTB-31 [C-33A]; Manassas, VA, USA).

Techniques: Activity Assay, MTT Assay

Journal: International Journal of Molecular Sciences

Article Title: Reactivation of P53 Antiproliferative and Pro-Apoptotic Pathways by Resveratrol in Mutant P53 Cancer Cell Lines

doi: 10.3390/ijms27104481

Figure Lengend Snippet: Resveratrol induces apoptosis in cancer cell lines with mutant or null p53 . ( A ) C33A, SK-BR-3, and Saos-2 cells were treated with vehicle (ethanol) or resveratrol at their respective IC50s for 48 h. Apoptosis was assessed by means of Annexin V/propidium iodide (PI) double staining followed by flow cytometry analysis. Representative dot plots show viable cells in the lower left quadrant (Annexin V − /PI − ), early apoptotic cells in the lower right quadrant (Annexin V + /PI − ), and late apoptotic cells in the upper right quadrant (Annexin V + /PI + ). ( B ) Quantifications of total apoptotic cells (early + late apoptosis) are shown as the mean ± SD from three independent experiments. ** p < 0.01; *** p < 0.001 versus EtOH. RES, resveratrol; EtOH, control with ethanol.

Article Snippet: The human cervical cancer cell line C33A ( p53 mutation R273C) was purchased from the American Type Culture Collection (ATCC cat. no. HTB-31 [C-33A]; Manassas, VA, USA).

Techniques: Mutagenesis, Double Staining, Flow Cytometry, Control

Journal: International Journal of Molecular Sciences

Article Title: Reactivation of P53 Antiproliferative and Pro-Apoptotic Pathways by Resveratrol in Mutant P53 Cancer Cell Lines

doi: 10.3390/ijms27104481

Figure Lengend Snippet: Resveratrol induces the expression of p53 target genes across different p53 backgrounds. Gene expression analysis was performed in ( A ) C33A ( p53 R273C), ( B ) SK-BR-3 ( p53 R175H), ( C ) MCF-7 ( p53 WT), and ( D ) Saos-2 ( p53 null) cells following treatment with vehicle (EtOH) or resveratrol (RES) at their respective IC50s for 48 h. Relative mRNA levels of p53 target genes were determined by means of RT-qPCR and expressed as fold changes relative to vehicle-treated controls, normalized to the housekeeping gene β2-microglobulin (β2M). Data are presented as the mean ± SD of at least three independent experiments. Statistical significance was evaluated using one-way ANOVA followed by appropriate post hoc multiple comparison tests. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001 versus control.

Article Snippet: The human cervical cancer cell line C33A ( p53 mutation R273C) was purchased from the American Type Culture Collection (ATCC cat. no. HTB-31 [C-33A]; Manassas, VA, USA).

Techniques: Expressing, Gene Expression, Quantitative RT-PCR, Comparison, Control

Journal: International Journal of Molecular Sciences

Article Title: Reactivation of P53 Antiproliferative and Pro-Apoptotic Pathways by Resveratrol in Mutant P53 Cancer Cell Lines

doi: 10.3390/ijms27104481

Figure Lengend Snippet: Resveratrol enhances cisplatin sensitivity in cancer cell lines with mutant or null p53 . ( A ) C33A, ( B ) SK-BR-3, and ( C ) Saos-2 cells were treated with increasing concentrations of cisplatin (CDDP; 2.5–20 μM) in the presence or absence of resveratrol (RES), at the corresponding IC 50 for each cell line, for 48 h. Cellular metabolic activity was assessed via MTT assay and normalized to that of vehicle-treated controls. Data represent the mean ± SD of at least four independent experiments. ( D ) IC50 values for CDDP were calculated using non-linear regression analysis (log [CDDP] vs. normalized response, variable slope model) using GraphPad Prism 6.01. * Previously published results from our working group.

Article Snippet: The human cervical cancer cell line C33A ( p53 mutation R273C) was purchased from the American Type Culture Collection (ATCC cat. no. HTB-31 [C-33A]; Manassas, VA, USA).

Techniques: Mutagenesis, Activity Assay, MTT Assay