Journal: Oncotarget

Article Title: Evaluation of an Actinomycin D/VX-680 aurora kinase inhibitor combination in p53-based cyclotherapy

doi:

Figure Lengend Snippet: (A) Pretreatment with low dose ActD decreases the proportion of 8N cells caused by VX-680 in NHDF cells. ActD was added 24 hours post-seeding. VX-680 was added 48 hours post-seeding. Cells were harvested 96 hours after seeding. (B) NHDFs were treated with 4 nM ActD for 72 hours, fixed and stained with Giemsa (top panel). In the bottom panel, ActD was removed from the medium and cells were allowed to proliferate for 6 days before Giemsa staining.

Article Snippet: NHDF cells were bought from Promocell.

Techniques: Staining

Journal: Oncotarget

Article Title: Evaluation of an Actinomycin D/VX-680 aurora kinase inhibitor combination in p53-based cyclotherapy

doi:

Figure Lengend Snippet: (A and B) NHDF cells were treated for 48 hours with VX-680 alone (labelled as no preinc.), or treated for 48 hours with VX-680 after preincubating with the indicated amounts of ActD for 24 hours. Thereafter cells were fixed, stained Giemsa (A) and counted (B). Arrows in (A) indicate cells with abnormal nuclei. (C and D) NHDF cells treated as in A and B were allowed to recover for 6 days in drug-free medium. The growth of the cultures in drug-free medium is quantified in panel (C) by counting the total number of cells per field before and after the recovery period. The average number of cells per field with aberrant nuclei before and after the recovery period is shown in panel (D). Error bars correspond to standard deviations.

Article Snippet: NHDF cells were bought from Promocell.

Techniques: Staining

Journal: Oncotarget

Article Title: Evaluation of an Actinomycin D/VX-680 aurora kinase inhibitor combination in p53-based cyclotherapy

doi:

Figure Lengend Snippet: (A) NHDF cells were treated with the indicated amounts of ActD for 4 or 12 hours. p53 and p21 were analysed by Western blotting. α-tubulin was detected as a loading control. (B) Example of nuclear abnormalities in NHDF cells expressing a dominant negative form of p53. The percentage of cells with this sort of aberrant nuclei was 2.5 fold higher than in the corresponding cells with intact p53. Severely damaged cells, such as the one in the center of this picture, only appeared in the fibroblasts with dominant negative p53.

Article Snippet: NHDF cells were bought from Promocell.

Techniques: Western Blot, Expressing, Dominant Negative Mutation

Journal:

Article Title: ?-Melanocyte-Stimulating Hormone Counteracts the Suppressive Effect of UVB on Nrf2 and Nrf-Dependent Gene Expression in Human Skin

doi: 10.1210/en.2008-1315

Figure Lengend Snippet: Expression analysis of Nrf1-3 in human skin cells in vitro and in situ. A, Detection of Nrf1-3 in various human skin cell types in vitro as shown by conventional RT-PCR analysis. NC, Negative control, H2O as template; 1, NHK; 2, HaCaT; 3, NHM; 4, HDF; 5, HMEC-1. B, Protein expression of Nrf2 in human skin cell types. Total cell lysates (50 μg/lane) were subjected to Western immunoblot analysis using an anti-Nrf2 antibody. The same blot was reprobed with an anti-α-tubulin antibody. C, Expression of Nrf2 in healthy human skin. Nrf2 was detected by immunohistochemistry using the immunoperoxidase technique (a). Note specific cytoplasmic Nrf2 immunoreactivity within the basal layer of epidermal keratinocytes (red cells, arrows) but not in the IgG control (c). Double immunostaining of Nrf2 and melanocytes by the immunoperoxidase (Nrf2) and immunogold-silver enhancement technique (melanocytes) using a pan-Mel antibody (b). Note gray-blackish precipitate within a melanocyte but lack of Nrf2 staining within (arrow).

Article Snippet: Cell culture Normal human keratinocytes (NHK) and human dermal fibroblasts (HDF) were all derived from newborn foreskin and purchased from PromoCell (Heidelberg, Germany).

Techniques: Expressing, In Vitro, In Situ, Reverse Transcription Polymerase Chain Reaction, Negative Control, Western Blot, Immunohistochemistry, Double Immunostaining, Staining