Fig. 5.

Mutation of TIEG1 Ser 384 and Thr 445 to glutamate induces subnuclear relocalization. a The phospho-mimicking mutants of TIEG1 are enriched in round nuclear bodies. HeLa and/or U2OS cells were transfected with an expression construct for EGFP-TIEG1 or EGFP-tagged TIEG1 mutants and imaged by confocal laser fluorescence microscopy 24 h post transfection. b The TIEG1 mutants display mobility similar to the wild-type TIEG1. HeLa cells were transfected with the indicated constructs, and 24 h post transfection the cells were imaged and fluorescence signals were collected before and after photobleaching a portion of the nucleus. The recovery kinetics were calculated after double normalization of the initial fluorescence, corrected for background and loss of fluorescence caused by imaging. The graph presented here shows the average intensity of the bleach spot of 9–10 independent cells for the indicated time points. Error bars represent the SD values between various experiments