Figures 3.

Stable transfection of HeLa cells was performed with gene optimized MR. Stables were selected with 1 mg/ml of G418 sulfate. The cells were incubated under selection pressure for several weeks with change of media and addition of G418 sulfate every other day. Following selection, cells were harvested and stained with monoclonal anti-MR and secondary stained with goat anti-mouse Alexa 488 or 555. Nuclear staining is achieved with ToPro-3. Images were captured with a 63× oil objective on the Zeiss LSM-510 (A) Demonstrates the cytometric profile of MR and TLR2 in HeLa cells stably expressing OptMR after in 1mg/ml G418. Shaded histogram represents the unstained control, TLR2 expression in blue histogram and MR expression in the red histogram. (B) Demonstrates the distribution of MR within stable transfectants. (C) Western blot analysis was performed on cell lysate from stable transfectants.