A simple and dual expression plasmid system in prokaryotic (<i>E</i>. <i>coli</i>) and mammalian cells - Fig 3

A. Transfection of pgMAX/DsRed2 (mammalian mode). Transfected plasmids are indicated. Phase-contrast (upper panels) and DsRed2 fluorescence (lower panels). pgMAX/DsRed exhibited strong red fluorescence comparable with DsRed2. pTarget/DsRed showed limited red fluorescence. B. Time-dependent changes in red fluorescence. Statistical analysis of red fluorescence of DsRed2-transfected cells (open box), pgMAX/DsRed-transfected cells (filled circle) (n = 8), pTarget/DsRed-transfected cells (open triangle) (n = 8), and pgMAX-transfected cells (negative control; open circle) (n = 8). Fluorescence of DsRed2 and pgMAX/DsRed increased up to the third day (3 days after transfection) (*p #p C. Comparative RT-PCR analysis of DsRed2. Comparative RT-PCR with DsRed2 specific primers resulted in comparable expression of DsRed2 with DsRed2 (D) and pgMAX/DsRed (M). A negative control (N), HEK293T cells (non-transfected) and DNA ladder (L) are shown. D. Western blot analysis using anti-flag, anti-DsRed and anti-GAPDH antibodies. The anti-flag antibody recognized a 33-kDa band in pgMAX/DsRed-transfected cells (M). The anti-DsRed antibody recognized a 30-kDa band in DsRed2-transfected cells (D) and a 33-kDa band in pgMAX/DsRed-transfected cells (M). The negative control (N), HEK293T cells (non-transfected), is shown. GAPDH was used as an endogenous control.