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. Author manuscript; available in PMC: 2012 Aug 19.
Published in final edited form as: Exp Cell Res. 2007 Jul 19;313(15):3261–3275. doi: 10.1016/j.yexcr.2007.07.009

Fig. 4.

Fig. 4

Interaction of ectopically expressed HA-Erk2 and HA-Sgk to recombinant full length and truncated importin-alpha. Panel A: GST-FLIa (full length importin-alpha) fusion protein is composed of a GST epitope tag, amino-terminal domain (NTD), Arm repeat containing domain, and a carboxy-terminal domain (CTD). GST-Tia (truncated importin-alpha) is composed of GST linked to the carboxy-terminal 106 amino acids (amino acids 423-529). GST alone was used as a negative control. Panel B: Hek-293 cells were transiently transfected with expression plasmids encoding either wild-type HA epitope tagged Erk2 (HA-Erk2) or Sgk (WT-HA-Sgk) or HA-Jnk or vector alone, and 48 hours post-transfection cell lysates were prepared as described in the Methods section. GST protein alone or GST-FLIa or GST-TIa was incubated with the indicated lysates containing exogenously expressed wild-type HA-tagged Sgk, or HA-Erk2, or HA-Jnk. The proteins bound to the Glutathione bead-bound GST or GST-fusion proteins were separated on SDS-PAGE and immunoblotted with anti-HA antibodies to detect bound proteins. Inputs denote 10% of the extract and lysates prepared from vector transfected samples served as controls as shown in each panel. Molecular weight markers are shown on the left side of each panel. Experiments were repeated three times with similar results.