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. Author manuscript; available in PMC: 2015 Nov 20.
Published in final edited form as: Chem Biol. 2014 Oct 23;21(11):1564–1574. doi: 10.1016/j.chembiol.2014.09.009

Figure 1. Stress-Independent ATF6 Preactivation Preferentially Decreases Secretion of Disease-Associated TTR Variants.

Figure 1

(A) Schematic illustrating the TMP-mediated, posttranslational regulation of DHFR.ATF6 in HEK293DAX cells (Shoulders et al., 2013).

(B–H) Quantification of fraction secreted [35S]-labeled FTTTR variants from HEK293DAX cells in the absence (black) or presence (blue) of ATF6 preactivation (TMP, 10 μM; 16 hr). Representative autoradiograms and the experimental protocol are shown in Figure S1A. Fraction secreted was calculated from autoradiograms using the equation: Fraction Secreted = {Total FTTTR Signal in Media at Time = t } divided by {Total FTTTR Signal in Lysate at Time = 0} + {Total FTTTR Signal in Media at Time = 0} (Sekijima et al., 2005; Shoulders et al., 2013). Error bars represent SEM from biological replicates (n ≥ 3). Identical plots of fraction secreted for FTTTRD18G and FTTTRA25T with the same scales used for the other TTR variants are shown in Figures S1B and S1C.

(I) Bar graph comparing fraction secreted at 4 hr for FTTTR variants in the absence (gray) or presence of ATF6 preactivation (blue), as shown in Figures 1B–1H. Shading shows familial disease-associated FTTTR variants (pink) and FTTTR variants not involved in familial disease (brown). Error bars represent SEM from biological replicates (n ≥ 3).

**p < 0.01; ***p < 0.001.