An Inducible Expression System of the Calcium-Activated Potassium
Channel 4 to Study the Differential Impact on Embryonic Stem Cells
Figure 3
Characterization of iSK4 cells. (a) Flow cytometry by FACS shows a major proportion of the doxycycline-treated cells to exhibit green fluorescence (>75%). (b, c) Neomycin-selected iSK4 clonal cultures show dox-dependent elevation of SK4 proteins. (b) Iimmunocytochemistry using specific SK4 antibodies shows highly elevated protein levels of SK4 in the colonies after 1 day of doxycycline treatment. Dox-treated cells exhibit the AcGFP fluorescence (green) in contrast to controls. Both treated and untreated cells express SK4 proteins (red) with a strong increase in the dox-treated cells. Elevated SK4 proteins upon dox treatment was underlined by (c) by immunoblot. Nuclei are stained by DAPI (blue). Scale bars as indicated. (d) SK4 transcript increase upon doxycycline treatment is validated by real-time one-step RT-PCR.
