Journal: Oncogene

Article Title: p21(Waf1/Cip1) protects against p53-mediated apoptosis of human melanoma cells.

doi: 10.1038/sj.onc.1200897

Figure Lengend Snippet: Figure 1 p53 overexpression induces apoptosis of SK-MEL-110 (MEL) cells. (a) Eect of Ad.null or AdCMV.p53 infections in SK-MEL-110 cells and VSMC. Infected cells (100 p.f.u./cell in all experiments) were analysed for relative changes in cell number (counted using a hemocytometer) 48 h after infection with the indicated viruses (solid bars, Ad.null; hatched bars, AdCMV.p53). Values represent mean+SEM of three independent experiments. (b) Apoptotic cell death of SK-MEL-110 cells infected with AdCMV.p53. Thirty-six hours after infection of VSMC and SK- MEL-110 cells with 100 p.f.u./cell Ad.null (null) or AdCMV.p53 (p53), genomic DNA was extracted and assayed from fragmenta- tion. MW: Molecular weight marker

Article Snippet: Fifty-mg samples of total cell lysates were size-fractionated by SDS ± PAGE and transferred onto PVDF membranes using standard techniques (Harlow and Lane, 1988). p53 and p21Waf1/Cip1 proteins were detected with the ECL system (Amersham, Arlington Heights, IL) following incubation with the monoclonal anti-human p53 antibody (Santa Cruz Biotechnology, Santa Cruz, CA) and rabbit polyclonal anti-p21 antibody (Oncogene Science).

Techniques: Over Expression, Infection, Molecular Weight, Marker

Journal: Oncogene

Article Title: p21(Waf1/Cip1) protects against p53-mediated apoptosis of human melanoma cells.

doi: 10.1038/sj.onc.1200897

Figure Lengend Snippet: Figure 2 (a) p53 expression in VSMC and SK-MEL-110 cells. Cells were infected with 100 p.f.u./cell AdCMV.p53 and p53 expression was monitored 24 h later by Western blot analysis. (b) Northern blot analysis of p21Waf1/Cip1, GADD45 and p53 mRNA expression in Ad.null- or AdCMV.p53-infected VSMC and SK-MEL-110 cells. Expression was monitored at the times indicated following infection. Loading and transfer dierences among samples were normalized using an oligomer recognizing 18S rRNA

Article Snippet: Fifty-mg samples of total cell lysates were size-fractionated by SDS ± PAGE and transferred onto PVDF membranes using standard techniques (Harlow and Lane, 1988). p53 and p21Waf1/Cip1 proteins were detected with the ECL system (Amersham, Arlington Heights, IL) following incubation with the monoclonal anti-human p53 antibody (Santa Cruz Biotechnology, Santa Cruz, CA) and rabbit polyclonal anti-p21 antibody (Oncogene Science).

Techniques: Expressing, Infection, Western Blot, Northern Blot

Journal: Oncogene

Article Title: p21(Waf1/Cip1) protects against p53-mediated apoptosis of human melanoma cells.

doi: 10.1038/sj.onc.1200897

Figure Lengend Snippet: Figure 3 (a) p21Waf1/Cip1 protein expression in SK-MEL-110 cells 24 h after infection with the indicated viruses. (b) Loss in cell viability of control SK-MEL-110 cultures infected with AdCMV.p53 relative to that of p21Waf1/Cip1-overexpressing cells infected with AdCMV.p53. Abscissa, ®rst infection with Ad.null (left), or AdWAF1 (right); second infection: solid bars (Ad.null), hatched bars (AdCMV.p53). Values represent mean+SEM of at least three independent experiments. (c) Time-dependent changes in cell number of SK-MEL-110 cultures that had been infected with Ad.null (null) or AdWAF1 (p21) 48 h prior to a subsequent infection with either Ad.null or AdCMV.p53 (p53). Infection groups: null/null (- -&- -), null/p53 (±&±), p21/null (- -&- -) and p21/p53 (±&±). Values represent mean+SEM of at least three independent experiments. (d) FACS analysis of SK-MEL-110 cultures after sequential infections with Ad.null (null) or AdWAF1 (p21) for 48 h (®rst infection) followed by Ad.null (null) or AdCMV.p53 (p53) for the following 48 h (second infection). Note the apoptotic peak (arrowhead), readily visible in the null/p53 treatment group

Article Snippet: Fifty-mg samples of total cell lysates were size-fractionated by SDS ± PAGE and transferred onto PVDF membranes using standard techniques (Harlow and Lane, 1988). p53 and p21Waf1/Cip1 proteins were detected with the ECL system (Amersham, Arlington Heights, IL) following incubation with the monoclonal anti-human p53 antibody (Santa Cruz Biotechnology, Santa Cruz, CA) and rabbit polyclonal anti-p21 antibody (Oncogene Science).

Techniques: Expressing, Infection, Control

Journal: Oncogene

Article Title: p21(Waf1/Cip1) protects against p53-mediated apoptosis of human melanoma cells.

doi: 10.1038/sj.onc.1200897

Figure Lengend Snippet: Figure 4 p53-mediated cytotoxicity in mouse embryonal ®broblasts lacking the p21Waf1/Cip1 gene (p217/7 MEFs) is prevented by prior expression of p21Waf1/Cip1. (a) Western blot analysis of p53 expression in p217/7 MEFs following infection with either Ad.null AdCMV.NLSbgal or AdCMV.p53 adenoviruses at the indicated p.f.u./cell. (b) Cytotoxic eect of p53 overexpression in p217/7

Article Snippet: Fifty-mg samples of total cell lysates were size-fractionated by SDS ± PAGE and transferred onto PVDF membranes using standard techniques (Harlow and Lane, 1988). p53 and p21Waf1/Cip1 proteins were detected with the ECL system (Amersham, Arlington Heights, IL) following incubation with the monoclonal anti-human p53 antibody (Santa Cruz Biotechnology, Santa Cruz, CA) and rabbit polyclonal anti-p21 antibody (Oncogene Science).

Techniques: Expressing, Western Blot, Infection, Over Expression