Fig. 5.

Gel shift assays were performed for nuclear factor of activated T cells (NFAT) and myocyte enhancing factor 2 (MEF-2) and quantified using the Licor IR probe sets. With social stress, there was a rise in the nuclear translocation of both NFAT and MEF-2 over that seen in controls. In the absence of nuclear protein fractions, signal was lost (lane 1) compared with the gel shift seen from nuclear proteins from a social stress bladder (lane 2). Signal intensity for both NFAT and MEF-2 was lost with the application of “cold” oligonucleotide probe to the reaction (lane 3). The specificity of the oligonucleotide probe for the transcription factors was also studied using monocloncal antibodies. With the addition of anti-NFATc3 mab, there was a loss of signal as previously described (lane 4). A similar loss of signal was seen with antibodies against NFATc4 (data not shown). With addition of the monoclonal antibody against the MEF-2b, a pronounced supershift (SS) was observed (lane 4). These values represent the average ± SD from 5 control and 5 stressed mice.