Journal:

Article Title: Disruption of the nucleolus mediates stabilization of p53 in response to DNA damage and other stresses

doi: 10.1093/emboj/cdg579

Figure Lengend Snippet: Fig. 1. Nucleolar disruption is independent of p53. (A–G′) Single confocal sections of nuclei of NDFs (A–G) and 041 cells (A′–G′) stained for fibrillarin, 6 h after treatment with each indicated agent. Each image corresponds to a single nucleus. (A′′–G′′′) wide field images of NDFs (A′′–G′′) and 041 cells (A′′′–G′′′) stained for NPM after the same treatments. A pseudo-colour scale (indicated) was applied to each image to highlight all intensity ranges. (H) Dual plot of nuclear p53 expression level (DO-1 staining) and NPM translocation index for NDFs treated for 6 h with the indicated agents.

Article Snippet: Membranes were probed with either DO-1 antibody, anti-p53 Ser15-P rabbit polyclonal (Santa Cruz) or anti-p53 Ser392-P mouse monoclonal antibodies (AutogenBioclear, Calne, UK), followed by horseradish peroxidase (HRP)-labelled secondary antibody (Dako, Ely, UK).

Techniques: Staining, Expressing, Translocation Assay

Journal:

Article Title: Disruption of the nucleolus mediates stabilization of p53 in response to DNA damage and other stresses

doi: 10.1093/emboj/cdg579

Figure Lengend Snippet: Fig. 2. Effects of micropore irradiation on p53 expression and NPM translocation in NDFs. (A) Distribution of the fraction of irradiated areas on NDF nuclei (n = 1699) observed by the ratio between the area exposed to UV irradiation through 3 µm Isopore filters (irradiated area detected by antibody labelling of photolesions) and the total nuclear projected area (Hoechst 33324). The insert shows an example field of micropore-irradiated nuclei, with CPDs labelled red and nuclei blue (Hoechst). (B) p53 expression levels in NDFs whole-nucleus irradiated at 10 J/m2 (red), micropore irradiated at 40 (green), 60 (yellow) and 80 J/m2 (blue), and non-irradiated (black). The fractions of nuclei receiving WED ≥10 J/m2 (see text) under each micropore irradiation condition are indicated in (A). (C) NPM translocation indext for NDFs irradiated in the same conditions as in (B). Cells in (A) were fixed immediately after irradiation, while cells in (B) and (C) were fixed 6 h after irradiation.

Article Snippet: Membranes were probed with either DO-1 antibody, anti-p53 Ser15-P rabbit polyclonal (Santa Cruz) or anti-p53 Ser392-P mouse monoclonal antibodies (AutogenBioclear, Calne, UK), followed by horseradish peroxidase (HRP)-labelled secondary antibody (Dako, Ely, UK).

Techniques: Irradiation, Expressing, Translocation Assay

Journal:

Article Title: Disruption of the nucleolus mediates stabilization of p53 in response to DNA damage and other stresses

doi: 10.1093/emboj/cdg579

Figure Lengend Snippet: Fig. 3. Correlation of MRD for p53 expression and NPM translocation in NDFs and Cockayne syndrome complementation group A (CS-A) fibroblasts. NDFs (A–D) and CS-A cells (E–H) were irradiated at the indicated UV densities, fixed 6 h later and stained for p53 expression (A–D and E–H, with positions of nuclei indicated by Hoechst staining in A′–D′ and E′–H′) or NPM (A′′–D′′ and E′′–H′′). All images are wide-field. Pseudo-colour is used in NPM images.

Article Snippet: Membranes were probed with either DO-1 antibody, anti-p53 Ser15-P rabbit polyclonal (Santa Cruz) or anti-p53 Ser392-P mouse monoclonal antibodies (AutogenBioclear, Calne, UK), followed by horseradish peroxidase (HRP)-labelled secondary antibody (Dako, Ely, UK).

Techniques: Expressing, Translocation Assay, Irradiation, Staining

Journal:

Article Title: Disruption of the nucleolus mediates stabilization of p53 in response to DNA damage and other stresses

doi: 10.1093/emboj/cdg579

Figure Lengend Snippet: Fig. 4. Effect of nucleolar disruption by microinjection of an anti-UBF antibody in NDFs. (A, C and E) Microinjection of anti-UBF (IgG1) with fibrillarin distribution (A) and p53 expression (C) assayed 6 h post-injection, and p21 expression (E) 18 h post-injection. (B, D and F) Microinjection of control purified mouse IgG1. (A′–F′) Reference Hoechst images. Arrows indicate microinjected cells. Lower magnification was used in p21 images in order to incorporate more cells into the field of view. (G–J) p53 phosphorylation in response to microinjection of anti-UBF (G and I) and non-specific IgG (H and J). p53 phosphorylation was detected for Ser15 (G and H) and Ser392 (I and J). (G′–J′) reference Hoechst images. (K–N) Phosphorylation at Ser15 and Ser392 in control and UV-irradiated NDFs (14 J/m2) as indicated. (O) Western blot analysis of p53 expression and phosphorylation (Ser15 and Ser392) in control and UV-irradiated NDFs (1–2, 4 and 14 J/m2). Cells were harvested 6 h post-irradiation.

Article Snippet: Membranes were probed with either DO-1 antibody, anti-p53 Ser15-P rabbit polyclonal (Santa Cruz) or anti-p53 Ser392-P mouse monoclonal antibodies (AutogenBioclear, Calne, UK), followed by horseradish peroxidase (HRP)-labelled secondary antibody (Dako, Ely, UK).

Techniques: Expressing, Injection, Purification, Irradiation, Western Blot

Journal: Biological Research

Article Title: CoCl 2 simulated hypoxia induce cell proliferation and alter the expression pattern of hypoxia associated genes involved in angiogenesis and apoptosis

doi: 10.1186/s40659-019-0221-z

Figure Lengend Snippet: Differential expression pattern of HIF-1α and hypoxia associated genes involved in angiogenesis and apoptosis like VEGF, p53 and BAX in breast cancer cells, where graphical data represents the relative density value after densitometry analysis of band of PCR amplified product. a Depicts the expression pattern of β-actin, HIF-1α and VEGF in MCF-7 cells treated with different concentration of CoCl 2 as compare to control by RT-PCR, similarly b shows expression of β-actin, HIF-1α, VEGF, p53 and BAX genes in MDA-MB-231 cells treated with increasing concentration of CoCl 2 , where it was clearly observed that expression of these genes increases significantly in a dose dependent manner. Error bar represents mean ± SEM of minimum three independent experiments and * Denotes significant difference (P < 0.05) as compared to control (N)

Article Snippet: Consequently, primary antibody for each protein like anti-β-actin (Mouse monoclonal antibody obtained from ambion by life technologies; Cat. No. AM4302), anti-p53 (Mouse monoclonal antibody from Puregene by Genetix Biotech Asia Pvt.

Techniques: Expressing, Amplification, Concentration Assay, Reverse Transcription Polymerase Chain Reaction

Journal: Biological Research

Article Title: CoCl 2 simulated hypoxia induce cell proliferation and alter the expression pattern of hypoxia associated genes involved in angiogenesis and apoptosis

doi: 10.1186/s40659-019-0221-z

Figure Lengend Snippet: Western blot analysis of β -actin, HIF-1α, VEGF, p53 and BAX protein in breast cancer cells grown under hypoxia (treated with CoCl 2 ) as well as in normoxia (without treatment). a Differential expression pattern of HIF-1α and VEGF protein isolated from MCF-7 cells. b Shows the dose dependent enhancement in expression of hypoxia associated proteins HIF-1α, VEGF, p53 and BAX in MDA-MB-231 cells grown under hypoxic condition, P < 0.05

Article Snippet: Consequently, primary antibody for each protein like anti-β-actin (Mouse monoclonal antibody obtained from ambion by life technologies; Cat. No. AM4302), anti-p53 (Mouse monoclonal antibody from Puregene by Genetix Biotech Asia Pvt.

Techniques: Western Blot, Expressing, Isolation

Journal: Biological Research

Article Title: CoCl 2 simulated hypoxia induce cell proliferation and alter the expression pattern of hypoxia associated genes involved in angiogenesis and apoptosis

doi: 10.1186/s40659-019-0221-z

Figure Lengend Snippet: Relative fold change in expression of HIF-1α and VEGF in MCF-7 cells treated with CoCl 2 in a dose dependent manner has been analysed in qPCR data a as compare to cells grown under normoxia, where β-actin used as a endogenous control. While b represents the qPCR data of HIF-1α, VEGF, p53 and BAX gene in MDA-MB-231 cells, P < 0.05

Article Snippet: Consequently, primary antibody for each protein like anti-β-actin (Mouse monoclonal antibody obtained from ambion by life technologies; Cat. No. AM4302), anti-p53 (Mouse monoclonal antibody from Puregene by Genetix Biotech Asia Pvt.

Techniques: Expressing

Journal: PLoS ONE

Article Title: Unambiguous Detection of Multiple TP53 Gene Mutations in AAN-Associated Urothelial Cancer in Belgium Using Laser Capture Microdissection

doi: 10.1371/journal.pone.0106301

Figure Lengend Snippet: Clinical, pathological and TP53 gene mutational data of areas with extensive p53 IHC overexpression in TCC of Belgian AAN patients.

Article Snippet: They were incubated overnight at room temperature with the anti-p53 mouse monoclonal antibody DO-7 (Biocarta, Europe Gmbh) at a dilution of 1∶1000.

Techniques: Over Expression

Journal: PLoS ONE

Article Title: Unambiguous Detection of Multiple TP53 Gene Mutations in AAN-Associated Urothelial Cancer in Belgium Using Laser Capture Microdissection

doi: 10.1371/journal.pone.0106301

Figure Lengend Snippet: A–G TCC area containing more than 60% p53 stained nuclei by IHC counterstained with hematoxylin from frozen section of ureter (A). For laser capture microdissection, a toluidine blue stained area in a representative frozen section was matched (B), cut (C) and catapulted (D) in the cap of a microtube (E). 3′-5′dideoxy sequencing electropherogram of the segment of p53 showing the wild type sequence (F, arrow) and A>T transversion (G, arrow) leading to missense mutation E286V in exon 8.

Article Snippet: They were incubated overnight at room temperature with the anti-p53 mouse monoclonal antibody DO-7 (Biocarta, Europe Gmbh) at a dilution of 1∶1000.

Techniques: Staining, Laser Capture Microdissection, Sequencing, Mutagenesis