Figure 7.

Granulins 2 and 3 promote TDP-43 accumulation. A, Intact DA/DB motor neurons (cell bodies and axons) were quantified for pgrn-1(-) and TDP-43;gran 3 animals expressing the P_unc-129::GFP neuronal marker. No significant differences were found (n = 10). B, VC neurons (cell bodies and axons) were counted for pgrn-1(-) and TDP-43;gran 3 animals expressing the P_lin-11::GFP neuronal marker. No significant differences were found (n = 10). C, Images of TDP-43::GFP nuclear localization in Day 1 adult worms. Arrows indicate neuronal nuclei. TDP-43 was observed only in nuclei and not in neuronal cell bodies or extensions. D, Western blot against total TDP-43 and phospho-TDP-43 (S409/S410) for indicated strains. E, Quantitation of total TDP-43 from Western blots, normalized to actin loading control. Data shown represents the average of three independent trials (unpaired Mann–Whitney test, single tail, *p < 0.05; error bars represent SEM). F, TDP-43 mRNA levels were determined by qPCR using two different primer sets for TDP-43. Shown is the fold-change in expression compared with the pgrn-1;TDP-43 strain. No significant differences were observed (n = 3 independent biological replicates, one-way ANOVA, error bars represent SD). G, Granulin does not affect deg-1(d)-induced touch cell necrosis. Necrotic PVC touch receptor neurons were quantified in strains through development and the percentage of animals demonstrating necrotic cells determined (n ≥ 120, error bars indicate SEM). H, Granulin does not further impair motor control in tau transgenic strains (n = 12; ns, not significant, one-way ANOVA; error bars represent SD).