Fig. 2

Ethosuximide acts independently of T-type calcium channels and bacterial metabolism to reduce Tau aggregation. a Ethosuximide Inhibition of the T-type calcium channel, CCA-1, is not required for protection against paralysis. Tau V337M transgenic animals were crossed with loss-of-function mutations for cca-1(ad1650) to generate homozygous cross progeny. Ethosuximide supplementation increased thrashing activity of the Tau V337M transgenic strain and the double mutant Tau V337M; cca-1(ad1650) strain to similar extents. Data are shown as mean ± SEM (**p < 0.01, *p < 0.05; n = 30-50 worms per data point). b Ethosuximide extends lifespan in Tau V337M mutants in the absence of CCA-1. Lifespan assays were performed on single mutant cca-1(ad1650), transgenic (Tau V337M) and double mutant Tau V337M; cca-1(ad1650) strains grown in the presence (dashed lines) or absence (solid lines) of 1 mg/ml ethosuximide (n > 100 worms per strain/condition). c Ethosuximide extends lifespan using killed bacteria as a food source. Lifespan assays were performed on Tau V337M worms grown under control conditions or in the presence of kanamycin, or 1 mg/ml ethosuximide or both (n > 80 worms per condition). d Total Tau protein expression in Tau V337M worm lysates is not reduced by ethosuximide. The left panel shows a representative western blot; the right panel shows quantification of Tau normalised to actin and expressed as % of untreated control (mean ± SEM, n = 3; not significant). e, f Ethosuximide affects Tau proteostasis. e shows a representative western blot of the soluble and detergent-soluble (RIPA) sequentially extracted fractions in the presence or absence of ethosuximide treatment. f shows quantification of Tau fractions normalised to actin and expressed as % of the total (soluble + RIPA) protein level (data are shown as mean ± SEM (n = 3; *p < 0.05)