Journal: PLOS One

Article Title: Characterization of p53 p.T253I as a pathogenic mutation underlying Li-Fraumeni Syndrome

doi: 10.1371/journal.pone.0320036

Figure Lengend Snippet: HEK293 p53 -/- cells and those complemented with WT-GFP, C176Y-GFP, R213X-GFP, or T253I-GFP p53 were subjected to subcellular fractionation to look at the distribution of p53 and MDM2 in the cytoplasm ( A, left side ) and nucleus ( A, right side ). (B) Graphical depictions of the cytoplasmic ( left ) and nuclear ( right ) expression levels of p53. (C) Graphical depictions of the cytoplasmic ( left ) and nuclear ( right ) expression levels of MDM2. In both cases, expression levels of cytoplasmic fractions were normalized to GAPDH while expression levels of nuclear fractions were normalized to Histone 3 (H3). Data represent the mean expression levels of three experimental replicates and significance is denoted by * or † (* indicates p < 0.05 when comparing the sample to p53 -/-, † p < 0.05 when comparing the sample to p53 WT).

Article Snippet: Cat. # 4331182) were used to determine p21 (Cdkn1a Gene Expression Assay ID Hs00355782_ml) and MDM2 (Gene Expression Assay ID: Hs99999008_m1) gene expression levels .

Techniques: Fractionation, Expressing

Journal: PLOS One

Article Title: Characterization of p53 p.T253I as a pathogenic mutation underlying Li-Fraumeni Syndrome

doi: 10.1371/journal.pone.0320036

Figure Lengend Snippet: HEK293 p53 -/- cells and those complemented with WT-GFP, C176Y-GFP, R213X-GFP, or T253I-GFP p53 were treated with 5 ug/mL Cisplatin (or a DMSO vehicle control) for one hour followed by a 2-hour recovery period. Cells were collected and a Western blot was run looking at p53 signaling events. Levels of p53, MDM2, ( A, left side ) and p21 ( A, right side ) were examined with or without damage in each of the p53 conditions (null, WT, and the three mutants). Graphical depictions of p53 (B) , p21 (C) , and MDM2 ( D ) are shown. Data represent the mean expression levels, normalized to GAPDH, of three experimental replicates and significance is denoted by * or † (* indicates p < 0.05 when comparing the sample to p53 -/-, † p < 0.05 when comparing the sample to p53 WT).

Article Snippet: Cat. # 4331182) were used to determine p21 (Cdkn1a Gene Expression Assay ID Hs00355782_ml) and MDM2 (Gene Expression Assay ID: Hs99999008_m1) gene expression levels .

Techniques: Control, Western Blot, Expressing

Journal: PLOS One

Article Title: Characterization of p53 p.T253I as a pathogenic mutation underlying Li-Fraumeni Syndrome

doi: 10.1371/journal.pone.0320036

Figure Lengend Snippet: HEK293 p53 -/- cells and those complemented with WT-GFP, C176Y-GFP, R213X-GFP, or T253I-GFP p53 were treated with 5 ug/mL Cisplatin (or a DMSO vehicle control) for one hour followed by a 2-hour recovery period. RNA was isolated from these cells and subjected to rtPCR in the presence of primers for p21 ( A ) and MDM2 (B) . Data represent the mean RNA expression levels of three experimental replicates and significance is denoted by * or † (* indicates p < 0.05 when comparing the sample to p53 -/-, † p < 0.05 when comparing the sample to p53 WT).

Article Snippet: Cat. # 4331182) were used to determine p21 (Cdkn1a Gene Expression Assay ID Hs00355782_ml) and MDM2 (Gene Expression Assay ID: Hs99999008_m1) gene expression levels .

Techniques: Control, Isolation, Reverse Transcription Polymerase Chain Reaction, RNA Expression