Journal: Nucleic Acids Research
Article Title: An engineered l-arginine sensor of Chlamydia pneumoniae enables arginine-adjustable transcription control in mammalian cells and mice
Figure Lengend Snippet: Impact of l -arginine and its derivatives on ART-controlled gene expression. ( A ) The structure of the l -arginine derivatives used in this study. ( B ) Sixty hours after co-transfection with pSH91 (P SV40 -ARG2-pA) and pSH93 (P ART1 -SEAP-pA), SEAP expression of CHO-K1 was measured. Cells were incubated in cell culture media containing 10 mg/l l -arginine and supplemented with 57.4, 574 or 5740 μM of l -arginine, l -ornithine, l -citrulline, agmatine, l -homoarginine, l -arginine methyl ester, l -arginine ethyl ester or l -canavanine (5740 μM correspond to 1 g/l l -arginine).
Article Snippet: Regulating l -arginine and l -arginine analogs l -arginine stock solution was prepared in water by adjusting 50 g/l l -arginine base (0.287 M, AppliChem, Darmstadt, Germany, Cat. No. A3653,0100) to pH 7.2 with HCl and used at the final concentrations indicated. l -ornithine hydrochloride (Fluka, Buchs, Switzerland, Cat. No. 75470) and l -citrulline (Fluka, Buchs, Switzerland, Cat. No. 27510), l -homoarginine hydrochloride (Acros Organics, Basel, Switzerland, Cat. No. 169090050) were dissolved in water (0.287 M) with pH adjusted to 7.2 by NaOH. l -arginine methyl ester dihydrochloride (Fluka, Buchs, Switzerland, Cat. No. 11030), l -arginine ethyl ester dihydrochloride (Sigma-Aldrich, Steinheim, Germany, Cat. No. A2883), agmatine sulfate (Sigma-Aldrich, Cat. No. A7127) and l -canavanine sulfate (Sigma-Aldrich, Cat. No. C9758) stock solutions were prepared by adjusting the pH of a 0.144 M solution to pH 7.2.
Techniques: Expressing, Cotransfection, Incubation, Cell Culture