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Detection of p16 expression and the SA-βGal assay reveals p16-dependent senescence. A. Immunohistochemistry was performed as per Lotfi et al, 1997 , using a polyclonal anti-p1 6 antibody (BD Pharmingen, San Diego, CA, USA) 24h post-infection. C6 cells were transduced with the parental pCL virus or with the pCLp16 virus. B. Western blot analysis of 293 cell lysate used as a positive control (lane 1), C6-pCL cell lysate (lane 2) or C6-pCLp16 cell lysate (lane 3). The <t>ECL-biotinylated</t> molecular weight standard (Mw, Amersham Pharmacia Biotech, Upsalla, Sweden) is included for orientation. Lysates from equal numbers of cells were prepared 24h post-infection. The p16 protein was concentrated in an immunoprecipitation step prior to electrophoresis and dectection with a rabbit polyclonal anti-p1 6 antibody (Santa Cruz Biotechnologies, Inc., Santa Cruz, CA, USA). C. The SA-βGal assay was used to detect senescence associated β-galactosidase activity in C6 cells transduced by either the pCL parental or the pCLp16 virus followed by selection for G418 resistance. Following selection, cells were fixed and stained with x-gal at pH 6.0. At this pH, only senescence associated β-galactosidase activity is detected.
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Detection of p16 expression and the SA-βGal assay reveals p16-dependent senescence. A. Immunohistochemistry was performed as per Lotfi et al, 1997 , using a polyclonal anti-p1 6 antibody (BD Pharmingen, San Diego, CA, USA) 24h post-infection. C6 cells were transduced with the parental pCL virus or with the pCLp16 virus. B. Western blot analysis of 293 cell lysate used as a positive control (lane 1), C6-pCL cell lysate (lane 2) or C6-pCLp16 cell lysate (lane 3). The <t>ECL-biotinylated</t> molecular weight standard (Mw, Amersham Pharmacia Biotech, Upsalla, Sweden) is included for orientation. Lysates from equal numbers of cells were prepared 24h post-infection. The p16 protein was concentrated in an immunoprecipitation step prior to electrophoresis and dectection with a rabbit polyclonal anti-p1 6 antibody (Santa Cruz Biotechnologies, Inc., Santa Cruz, CA, USA). C. The SA-βGal assay was used to detect senescence associated β-galactosidase activity in C6 cells transduced by either the pCL parental or the pCLp16 virus followed by selection for G418 resistance. Following selection, cells were fixed and stained with x-gal at pH 6.0. At this pH, only senescence associated β-galactosidase activity is detected.
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Detection of p16 expression and the SA-βGal assay reveals p16-dependent senescence. A. Immunohistochemistry was performed as per Lotfi et al, 1997 , using a polyclonal anti-p1 6 antibody (BD Pharmingen, San Diego, CA, USA) 24h post-infection. C6 cells were transduced with the parental pCL virus or with the pCLp16 virus. B. Western blot analysis of 293 cell lysate used as a positive control (lane 1), C6-pCL cell lysate (lane 2) or C6-pCLp16 cell lysate (lane 3). The <t>ECL-biotinylated</t> molecular weight standard (Mw, Amersham Pharmacia Biotech, Upsalla, Sweden) is included for orientation. Lysates from equal numbers of cells were prepared 24h post-infection. The p16 protein was concentrated in an immunoprecipitation step prior to electrophoresis and dectection with a rabbit polyclonal anti-p1 6 antibody (Santa Cruz Biotechnologies, Inc., Santa Cruz, CA, USA). C. The SA-βGal assay was used to detect senescence associated β-galactosidase activity in C6 cells transduced by either the pCL parental or the pCLp16 virus followed by selection for G418 resistance. Following selection, cells were fixed and stained with x-gal at pH 6.0. At this pH, only senescence associated β-galactosidase activity is detected.
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Detection of p16 expression and the SA-βGal assay reveals p16-dependent senescence. A. Immunohistochemistry was performed as per Lotfi et al, 1997 , using a polyclonal anti-p1 6 antibody (BD Pharmingen, San Diego, CA, USA) 24h post-infection. C6 cells were transduced with the parental pCL virus or with the pCLp16 virus. B. Western blot analysis of 293 cell lysate used as a positive control (lane 1), C6-pCL cell lysate (lane 2) or C6-pCLp16 cell lysate (lane 3). The <t>ECL-biotinylated</t> molecular weight standard (Mw, Amersham Pharmacia Biotech, Upsalla, Sweden) is included for orientation. Lysates from equal numbers of cells were prepared 24h post-infection. The p16 protein was concentrated in an immunoprecipitation step prior to electrophoresis and dectection with a rabbit polyclonal anti-p1 6 antibody (Santa Cruz Biotechnologies, Inc., Santa Cruz, CA, USA). C. The SA-βGal assay was used to detect senescence associated β-galactosidase activity in C6 cells transduced by either the pCL parental or the pCLp16 virus followed by selection for G418 resistance. Following selection, cells were fixed and stained with x-gal at pH 6.0. At this pH, only senescence associated β-galactosidase activity is detected.
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Detection of p16 expression and the SA-βGal assay reveals p16-dependent senescence. A. Immunohistochemistry was performed as per Lotfi et al, 1997 , using a polyclonal anti-p1 6 antibody (BD Pharmingen, San Diego, CA, USA) 24h post-infection. C6 cells were transduced with the parental pCL virus or with the pCLp16 virus. B. Western blot analysis of 293 cell lysate used as a positive control (lane 1), C6-pCL cell lysate (lane 2) or C6-pCLp16 cell lysate (lane 3). The <t>ECL-biotinylated</t> molecular weight standard (Mw, Amersham Pharmacia Biotech, Upsalla, Sweden) is included for orientation. Lysates from equal numbers of cells were prepared 24h post-infection. The p16 protein was concentrated in an immunoprecipitation step prior to electrophoresis and dectection with a rabbit polyclonal anti-p1 6 antibody (Santa Cruz Biotechnologies, Inc., Santa Cruz, CA, USA). C. The SA-βGal assay was used to detect senescence associated β-galactosidase activity in C6 cells transduced by either the pCL parental or the pCLp16 virus followed by selection for G418 resistance. Following selection, cells were fixed and stained with x-gal at pH 6.0. At this pH, only senescence associated β-galactosidase activity is detected.
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Image Search Results


Detection of p16 expression and the SA-βGal assay reveals p16-dependent senescence. A. Immunohistochemistry was performed as per Lotfi et al, 1997 , using a polyclonal anti-p1 6 antibody (BD Pharmingen, San Diego, CA, USA) 24h post-infection. C6 cells were transduced with the parental pCL virus or with the pCLp16 virus. B. Western blot analysis of 293 cell lysate used as a positive control (lane 1), C6-pCL cell lysate (lane 2) or C6-pCLp16 cell lysate (lane 3). The ECL-biotinylated molecular weight standard (Mw, Amersham Pharmacia Biotech, Upsalla, Sweden) is included for orientation. Lysates from equal numbers of cells were prepared 24h post-infection. The p16 protein was concentrated in an immunoprecipitation step prior to electrophoresis and dectection with a rabbit polyclonal anti-p1 6 antibody (Santa Cruz Biotechnologies, Inc., Santa Cruz, CA, USA). C. The SA-βGal assay was used to detect senescence associated β-galactosidase activity in C6 cells transduced by either the pCL parental or the pCLp16 virus followed by selection for G418 resistance. Following selection, cells were fixed and stained with x-gal at pH 6.0. At this pH, only senescence associated β-galactosidase activity is detected.

Journal: Cancer Cell International

Article Title: Retroviral transfer of the p16INK4a cDNA inhibits C6 glioma formation in Wistar rats

doi: 10.1186/1475-2867-2-2

Figure Lengend Snippet: Detection of p16 expression and the SA-βGal assay reveals p16-dependent senescence. A. Immunohistochemistry was performed as per Lotfi et al, 1997 , using a polyclonal anti-p1 6 antibody (BD Pharmingen, San Diego, CA, USA) 24h post-infection. C6 cells were transduced with the parental pCL virus or with the pCLp16 virus. B. Western blot analysis of 293 cell lysate used as a positive control (lane 1), C6-pCL cell lysate (lane 2) or C6-pCLp16 cell lysate (lane 3). The ECL-biotinylated molecular weight standard (Mw, Amersham Pharmacia Biotech, Upsalla, Sweden) is included for orientation. Lysates from equal numbers of cells were prepared 24h post-infection. The p16 protein was concentrated in an immunoprecipitation step prior to electrophoresis and dectection with a rabbit polyclonal anti-p1 6 antibody (Santa Cruz Biotechnologies, Inc., Santa Cruz, CA, USA). C. The SA-βGal assay was used to detect senescence associated β-galactosidase activity in C6 cells transduced by either the pCL parental or the pCLp16 virus followed by selection for G418 resistance. Following selection, cells were fixed and stained with x-gal at pH 6.0. At this pH, only senescence associated β-galactosidase activity is detected.

Article Snippet: The ECL biotinylated molecular size standard (Amersham Pharmacia Biotech, Sweden) was detected by HRP-streptavidin (Amersham Pharmacia Biotech, Upsalla, Sweden).

Techniques: Expressing, Immunohistochemistry, Infection, Transduction, Western Blot, Positive Control, Molecular Weight, Immunoprecipitation, Electrophoresis, Activity Assay, Selection, Staining