Characterization of the 4-OHT inducible cell lines. (A) Extracts from stable cell lines immunoblotted for HA and Tubulin (loading control). DAOY extracts (lane 1) show no expression of HA, while extracts from ER-MXD3.E66D (lane 2) and ER-MXD3 (lane 3) show an expected band of approximately 68 kDa. (B) Confocal images of the ER-MXD3 cell line treated with either ethanol (top panels) or 1 μM 4-OHT (bottom panels) for 1 hour and stained for DAPI, HA, and MXD3. When treated with 4-OHT the ER-MXD3 fusion protein is primarily localized in the nucleus (identified by DAPI staining) in contrast to vehicle (ethanol) treatment in which the fusion protein is localized throughout the cell. Scale bar 20 μm.
Ngo et al. BMC Cell Biology 2014 15:30 doi:10.1186/1471-2121-15-30