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Fig. 5
Heating or sonication restores lost trans-4-OHT potency. (A) mCherry intensity was quantified in embryos induced with >12 weeks old trans-4-OHT that was heated at 65°C or sonicated for 10 min prior to addition to E3 (scale bar 500 µm). (B) Heating or sonicating restores aged trans-4-OHT activity compared to both non-heated/non-sonicated compound to levels of the fresh compound (two-tailed, unpaired t-test, aged vs heated p = 0.0008, aged vs sonicated p = 0.0319, heated vs fresh p = 0.9029, sonicated vs fresh p = 0.1684). (C) EGFP-ERT2 cells were treated with different concentrations of control or heated (10 min at 65°C) aged trans-4-OHT. Images were taken 20 h post treatment and analyzed using Columbus software. Percentage of translocation was plotted using GraphPad Prism software. Heating moderately improves trans-4-OHT potency to induce translocation of EGFP-ERT2 to the nucleus (two-way ANOVA, p<0.0001).