Fig. 1

DCTN1 knockdown exacerbates TDP-43 toxicity in Drosophila. a Relative levels of dDCTN1 mRNA in adult fly brains analyzed by quantitative RT-PCR. Data were normalized using rp49 mRNA levels, and are presented as relative values to dDCTN1 mRNA levels of wild-type flies. Data are presented as the mean ± standard error of the mean (SEM) of three independent experiments. Each dot represents data from a single experiment. Fly genotypes: +, Elav-Gal4/+; dDCTN1-IR#1, Elav-Gal4/+;dDCTN1-IR#1/+; dDCTN1-IR#2, Elav-Gal4/dDCTN1-IR#2. b Light microscopic images of compound eyes of EGFP flies or TDP-43 flies coexpressing EGFP-IR or dDCTN1-IR (#1 or #2) under the control of the GMR-Gal4 driver. Arrows indicate necrotic patches. Scale bar, 200 μm. c Bar graphs showing the area of remaining eye pigment (left) and the area of necrotic patches (right) of TDP-43 flies in b. Each dot represents data from each fly (n = 24–26 flies). Fly genotypes: EGFP/EGFP-IR, GMR-Gal4/+;EGFP/+;EGFP-IR/+; EGFP/dDCTN1-IR#1, GMR-Gal4/+;EGFP/dDCTN1-IR#1; EGFP/dDCTN1-IR#2, GMR-Gal4/dDCTN1-IR#2;EGFP/+; TDP-43/EGFP-IR, GMR-Gal4/+;;TDP-43/EGFP-IR; TDP-43/dDCTN1-IR#1, GMR-Gal4/+;dDCTN1-IR#1/+;TDP-43/+; TDP-43/dDCTN1-IR#2, GMR-Gal4/dDCTN1-IR#2;;TDP-43/+. d Histological analysis of eye sections of adult flies. Scale bars, 50 µm (top) and 20 µm (bottom). Fly genotypes: +, GMR-Gal4/+; dDCTN1-IR, GMR-Gal4/+;dDCTN1-IR#1/+; TDP-43, GMR-Gal4/+;;TDP-43/+; TDP-43/dDCTN1-IR, GMR-Gal4/+;dDCTN1-IR#1/+;TDP-43/+. The bottom images are magnified images of the boxed regions. e Light microscopic images of compound eyes of EGFP flies or grim flies coexpressing EGFP-IR or dDCTN1-IR#1 under the control of the GMR-Gal4 driver. Fly genotypes: EGFP/EGFP-IR, GMR-Gal4/+;EGFP/+;EGFP-IR/+; EGFP/dDCTN1-IR, GMR-Gal4/+;EGFP/dDCTN1-IR#1; grim/EGFP-IR, GMR-Gal4/+;GMR-grim/+;EGFP-IR/+; grim/dDCTN1-IR#1, GMR-Gal4/+;GMR-grim/dDCTN1-IR#1. f Bar graph showing climbing scores of EGFP or TDP-43 flies coexpressing dDCTN1-IR in the nervous system at 7 days of age. Data are presented as the mean ± SEM of 4 to 6 independent experiments. Each dot represents data from a single experiment. Fly genotypes: EGFP, Elav-Gal4/+;EGFP/+; EGFP/dDCTN1-IR, Elav-Gal4/+;EGFP/dDCTN1-IR#1; TDP-43, Elav-Gal4/+;;TDP-43/+; TDP-43/dDCTN1-IR, Elav-Gal4/+;dDCTN1-IR#1/+;TDP-43/+. Statistical analyses in a, c, and f were performed to assess differences from control flies by one-way ANOVA followed by the Dunnett multiple comparison test (*p < 0.05, ***p < 0.001, ****p < 0.0001) (a, c), or to assess differences among groups by one-way ANOVA followed by the Tukey multiple comparison test (*p < 0.05, ***p < 0.001; n.s., not significant) (f)