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77955431877
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note
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m and Δ H u T m by fitting the fraction of monomer. Compound 2 was protonated by stirring with an excess of 2 M hydrochloric acid for 1 h. Compound 2-(6-HCl) was used.
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note
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The transcriptional activity of the p53 Arg337His mutant in the presence of calixarene was measured by a modified method as previously described in Ref. 4. NCI-H1299 cells (p53-null mutant cell line) were cultured in a 35 mm dish in RPMI-1640 medium containing 10% fetal calf serum. Cells were transfected with 0.5 μg of pEGFP-p53 and 3 μg of p53RE-mCherry reporter plasmid DNA with Lipofectamine 2000 (Invitrogen, USA) in OPTI-MEM. After 1 h, the medium was changed to RPMI-1640 medium with 10% fetal calf serum and calixarene derivatives were added to the medium. After 11 h incubation, cells were fixed with 3.5% formaldehyde, and EGFP (green) and mCherry (red) signals in the cells were quantified with a BZ-9000 (Keyence). Due to its poor solubility in medium, the transcriptional assay with compound 2 could not be performed.
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