Journal: Oncotarget

Article Title: Mitotic control of human papillomavirus genome-containing cells is regulated by the function of the PDZ-binding motif of the E6 oncoprotein

doi: 10.18632/oncotarget.14469

Figure Lengend Snippet: A . Abnormal (bar graph i) or normal (bar graph ii) mitoses were scored in each stage of mitosis (prophase, metaphase, anaphase and telophase) by α-tubulin staining and counterstaining with DAPI. The data are derived from three independent experiments for each of the three donors and presented as a proportion of total abnormal (top graph) or normal (bottom graph) mitoses. For each experiment, a minimum of 200 cells per donor was counted. B . Examples of abnormal mitoses in HPV18 genome-containing cells showing spindle morphology (α-tubulin, green) and chromosomes (DAPI, blue). C . Individual mitotic abnormalities were scored by α-tubulin staining and counterstaining with DAPI. Only the data for anaphase and telophase abnormalities are shown. The data are derived from three independent experiments for each of the three donors, with a minimum of 100 abnormal mitotic cells scored for each experiment, and the data is shown as the mean (± standard error of the mean). The differences in the frequency of individual abnormalities between WT and E6ΔPDZ cells were not statistically significant. D . The frequency of the combined late (anaphase and telophase) mitotic abnormalities in the E6ΔPDZ genome containing cells shown relative to cells harboring the WT genomes. Student's t test was used to determine the significance (*, P<0.05).

Article Snippet: Slides were then washed in a solution containing the DNA stain DAPI (Cytocell, Cambridge, United Kingdom) and for each cell passage, at least 200 cells with single nuclei visualized.

Techniques: Staining, Derivative Assay

Journal: Oncotarget

Article Title: Mitotic control of human papillomavirus genome-containing cells is regulated by the function of the PDZ-binding motif of the E6 oncoprotein

doi: 10.18632/oncotarget.14469

Figure Lengend Snippet: A . The proportion of cells containing micronuclei was scored from DAPI-stained cells. The data were collected from three independent experiments, with a minimum of 200 cells for each donor scored on each occasion. The data are shown as the mean (± standard error of the mean), and in all three donors the frequency was increased in E6ΔPDZ genome-containing cells (*, p<0.05); ns, not significant). B . Images of examples of CENPA positive and negative nuclei; in overlay CENPA (green), nuclei (blue). C . The proportion of cells with micronuclei containing centromeres was determined by immunofluorescence staining for CENPA. The data were collected from three independent experiments, with a minimum of 200 cells with micronuclei scored for each experiment, and shown as the mean (± standard error of the mean).

Article Snippet: Slides were then washed in a solution containing the DNA stain DAPI (Cytocell, Cambridge, United Kingdom) and for each cell passage, at least 200 cells with single nuclei visualized.

Techniques: Staining, Immunofluorescence

Journal: Oncotarget

Article Title: Mitotic control of human papillomavirus genome-containing cells is regulated by the function of the PDZ-binding motif of the E6 oncoprotein

doi: 10.18632/oncotarget.14469

Figure Lengend Snippet: Sections of organotypic raft cultures of A . untransfected HFK, B . HPV18 WT genome-containing cells and C . cells containing E6ΔPDZ genomes, were stained for α-tubulin (green), HPV18 E4 (red), and DNA (blue). Examples given in B and C were taken from multiple donors. The dotted white line in A, B and C indicates the junction between the basal cell layer and fibroblast embedded collagen matrix. D . The distribution of mitotic cells between the basal and suprabasal cell layers of organotypic rafts shown as a percentage of total α-tubulin stained mitotic cells (± standard error of the mean). Data were collected from three donors, and for each donor five sections of organotypic raft were analyzed (full thickness and entire length). Numbers of α-tubulin stained mitotic cells counted were 47 (HFK), 525 (WT) and 230 (E6ΔPDZ). Student's t test was used to determine significance (**, p<0.01). E . Sections were stained with γ-tubulin to visualize centrosomes and cells with visible centrosomes were scored for centrosome number. Rafts prepared from three different donors were analyzed; numbers of cells scored were: 1772 (HFK), 4022 (WT) and 2758 (E6ΔPDZ). Student's t test was used to determine significance (**, p<0.01; ***, p<0.001). Shown are examples of cells in E6ΔPDZ, WT rafts with >2 centrosomes, and an HFK raft with 2 centrosomes (red, γ-tubulin; blue, DAPI). The E6PDZ centrosomes are shown enlarged in the inset and the dotted white line indicates the junction between the basal cell layer and fibroblast embedded collagen matrix. Scale, 10 μm (inset 18.5 μm).

Article Snippet: Slides were then washed in a solution containing the DNA stain DAPI (Cytocell, Cambridge, United Kingdom) and for each cell passage, at least 200 cells with single nuclei visualized.

Techniques: Staining

Journal: FEBS Open Bio

Article Title: Angiotensin II type 1 receptor blocker telmisartan induces apoptosis and autophagy in adult T‐cell leukemia cells

doi: 10.1002/2211-5463.12055

Figure Lengend Snippet: Telmisartan causes G 1 phase cell cycle arrest. S1T, MT ‐2, Jurkat, and HL 60 cells were incubated in the absence or presence of telmisartan (100 μ m ) for 48 h, and then stained with DAPI and analyzed for DNA content by flow cytometry. Three independent experiments were performed per cell line; results are presented as mean percentages.

Article Snippet: Cell cycle analysis was performed with a DAPI Prep‐DNA staining solution (Sony).

Techniques: Incubation, Staining, Flow Cytometry