Fig. 5

Niclosamide ameliorates pathology in hFUS symptomatic mice. a Schematic illustration of niclosamide treatment in hFUS mice. Male mice were intraperitoneally injected daily with 20 mg/kg niclosamide from postnatal day (PND) 25 until death and spinal cord, sciatic nerves and skeletal muscles tissues were then analysed. b Niclosamide-treated hFUS mice (hFUS Nic) show a significant difference in the disease duration with respect to vehicle-treated hFUS mice (hFUS veh). Data are presented as means ± SEM. n = 6 mice/group. Two-tailed t test. t value: 4.719, degrees of freedom: 10. ***p < 0.001 vs. vehicle-treated hFUS mice. c Protein lysates from lumbar spinal cord of non-transgenic (Non-Tg) (~ 40 days), vehicle (hFUS veh) and niclosamide-treated hFUS mice (hFUS nic) at end stage of the disease were assayed by western blot with anti-GFAP, anti-S100A4, and anti-α-SMA. Data represent mean ± SEM of n = 4 mice/group. One-way ANOVA with Tukey correction between Non-Tg, hFUS veh and hFUS nic. F value (DFn, DFd): (2, 9) = 11.26 (GFAP), (2, 9) = 11.73 (S100A4), (2, 9) = 5.721 (a-SMA). *p < 0.05 and **p < 0.01 vs. Non-Tg mice or ^#p < 0.05 and ^##p < 0.01 vs. hFUS veh mice. d Representative fluorescence images of PDGFRβ (green) in the lumbar spinal cord of Non-Tg, hFUS veh, and hFUS nic mice at end stage of the disease. Scale bars: 50 μm. Immunofluorescence intensities were calculated by densitometric analyses. Data represent mean ± SEM. n = 4 mice/group, four sections per animal. One-way ANOVA with Tukey correction between Non-Tg, hFUS veh, and hFUS nic. F value (DFn, DFd): (2, 9) = 15.75 ***p < 0.001 vs. Non-Tg mice or ^#p < 0.05 vs. hFUS veh mice. e Representative fluorescence images of β-III tubulin (blue) and GFAP (purple) in the sciatic nerves of Non-Tg, hFUS veh, and hFUS nic mice at end stage of the disease. Scale bars: 50 μm. Immunofluorescence intensities were calculated by densitometric analyses. Data represent mean ± SEM. n = 4 mice/group, four sections per animal. One-way ANOVA with Tukey correction between Non-Tg, hFUS veh, and hFUS nic. F value (DFn, DFd): (2, 9) = 18.48 (β-III tubulin), (2, 9) = 17.94 (GFAP). *p < 0.05; **p < 0.01; and ***p < 0.001 vs. Non-Tg mice or ^#p < 0.05 and ^##p < 0.01 vs. hFUS veh mice. f Protein lysates from sciatic nerves of Non-Tg, hFUS veh, and hFUS Nic mice at end stage of the disease were assayed by western blot with anti-GFAP. GAPDH served as loading control. Relative densitometric values are reported on the right. Data represent mean ± SEM of n = 4 mice/group. One-way ANOVA with Tukey correction between Non-Tg, hFUS veh, and hFUS nic. F value (DFn, DFd): (2, 9) = 12.72. *p < 0.05 vs. Non-Tg mice or ^##p < 0.01 vs. hFUS veh mice