Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Human mesothelial cells are unusually susceptible to simian virus 40-mediated transformation and asbestos cocarcinogenicity.

doi: 10.1073/pnas.170207097

Figure Lengend Snippet: Fig. 3. Southern blot for SV40 DNA in HM3 and WI38. (A) SV40 DNA extracted from the cells. (B) SV40 DNA recovered in the tissue culture medium. Lanes: 1 and 2, HM 48 h after infection; 3 and 4, HF 48 h after infection; 5 and 6, HM 72 h after infection; 7 and 8, HF 72 h after infection. Each lane represents an independent infection experiment. Almost identical results were obtained with HM2 and MRC-5 HF. HM1 and CCD1069Sk were not tested. The different amounts of DNA (see text) were determined by Cherenkov counting of the individual lanes. DNAs were normalized for number of Tag-positive cells.

Article Snippet: 97 u no. 18 were used in our experiments at passages 17–18, and CCD1069Sk breast HF from a 70-year-old woman, which were used at passages 6–7, all from the American Type Culture Collection.

Techniques: Southern Blot, Infection

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Human mesothelial cells are unusually susceptible to simian virus 40-mediated transformation and asbestos cocarcinogenicity.

doi: 10.1073/pnas.170207097

Figure Lengend Snippet: Fig. 5. (A) p53 expression in HM and HF. One hundred micrograms of total protein extracts was loaded into each lane. GAPDH, glyceraldehyde-3-phosphate dehydrogenase. Lanes: 1, HM2; 2, HM3; 3, WI38; 4, CCD1069Sk [HM1 and MRC5 (not shown) produced almost identical results]. (B) Lanes 1–6, p53 expression in untreatedHM(lanes1and2),HMtreatedwith5mMscrambledoligo(lanes3and 4), and HM treated with 5 mM antisense p53 oligo (lanes 5 and 6). Cells were harvested and lysed 48 h after the onset of treatment; 100 mg of total protein extracts was loaded per lane (each lane represent an independent experiment). Lanes 7 and 8, p53 expression in HM 5 days after treatment (and 72 h after SV40 infection) with scrambled oligos (lane 7) and antisense p53 (lane 8). (C) Lanes 1 and 2, Tag immunoprecipitation in SV40-infected HM. Lane 1, scrambled control oligo-treated HM; lane 2, antisense p53-treated HM. Tag was precipitated with the monoclonal anti-Tag AB-1 (Tag amino terminus). The membrane was probed with the monoclonal anti-Tag AB-2 (Tag carboxyl terminus), followed by mono- clonal anti-mouse IgG conjugated with horseradish peroxidase. Lanes 3 and 4, Tagyp53 coimmunoprecipitation. The membrane shown on the left was stripped ofantibodiesandprobedwiththemonoclonalanti-p53DO-1directlyconjugated with horseradish peroxidase. Lane 3, control oligo-treated HM; lane 4, antisense p53-treated HM.

Article Snippet: 97 u no. 18 were used in our experiments at passages 17–18, and CCD1069Sk breast HF from a 70-year-old woman, which were used at passages 6–7, all from the American Type Culture Collection.

Techniques: Expressing, Produced, Infection, Immunoprecipitation, Control, Membrane