Figure 3
Higher temperature sensitivity of the Q25 TR-FRET signal is not due to other proteins or HTT protein level changes.
(a) Temperature sensitivity of the TR-FRET signals of Q25 versus Q72 HTT-exon1 proteins (125 ng/well) using 2B7/MW1, 4C9/MW1 and 2B7/4C9 TR-FRET antibody pairs. All signals (4°C (4C), room temperature (RT, 22°C) and 37°C (37C)) are normalized to the averaged signal at room temperature (RT) for each protein, n = 9, data plotted as average ± s.e.m. (b) 2B7/4C9 TR-FRET signals of the Q72, Q25 HTT-exon1 transfected samples or the non-transfected samples at 4°C (4°C), room temperature (RT), or 37°C (37C). (c) TR-FRET signals of the purified recombinant HTT-573 proteins with different polyQ length (Q25, Q46, Q75) at 4°C (4C) or room temperature (RT). Note that the temperature sensitivity is dependent on the polyQ length: Q25 > Q46 > Q72; n = 4, data plotted as average ± s.e.m. (d) TR-FRET signals of Q25 or Q72 transfected lysates, treated with or without RNase A. n = 3, data plotted as average ± s.e.m.
