Fig. 7

FICZ treatment of M17 cells increases endogenous TDP-43 stability. a Immunoblots of Avidin-agarose affinity purified (Avidin-AP) Click-iT AHA-labelled nascent proteins from lysates treated with DMSO vehicle or 0.5 μM FICZ, with a 2 h pulse of Click-iT AHA-labeling of endogenous nascent proteins then (by row) periods of culture in the absence of the Click-iT metabolite to observe degradation of AHA labelled nascent proteins. Blots are probed with anti-TARDBP antibody (see Additional file 3: Figure S4C). As negative controls for Avidin-AP, a representative sample from each time point, processed using the Click-iT chemistry but in the absence of the Biotin-alkyne chemoselective ligation tag, was also affinity purified against avidin-agarose (lanes 7: “No Biotin”). A further representative sample ligated using the Biotin-alkyne was affinity purified against control agarose resin as an additional negative control (lanes 8: “Agarose”). Densitometry of TDP-43 signal from dot blots (Additional file 3: Figure S4D) were quantified in b (n = 3; mean ± sd, curves fit by non-linear regression; t-tests were also performed for each data time point DMSO vs FICZ, ** P < 0.01, * P < 0.05). FICZ agonism of AHR increases the stability of TDP-43