Figure 2. Effect of physiological amino acid levels on transgenes expression.

Cells were cultivated on filters in standard cell culture medium and treated for the indicated times with physiological medium. A–B: mRNA expression of B⁰AT1 (A) or TMEM27 (B) was measured by quantitative PCR-analysis in B⁰AT1-TMEM27 overexpressing MDCK cells which were found mycoplasma-free (open bar) or mycoplasma-infected (black bar). mRNA levels were standardized to β-actin and normalized to time 0. C–D: Western blotting experiments with antibodies directed against B⁰AT1 (C) or TMEM27 (D) were performed in mycoplasma-infected B⁰AT1-TMEM27 overexpressing MDCK cells and the intensity of the immunoreactive bands was quantified, standardized to β-actin and normalized to time 0. Representative Western blotting images are shown. Data are represented as mean ± SEM (n = 3). Groups were compared by one-way ANOVA followed by Dunnett post-test; ** p≤0.01, * p≤0.05.