Fig. 6

Niclosamide reduces profibrotic and inflammatory pathways in the skeletal muscles of hFUS symptomatic mice. Protein lysates from gastrocnemius muscles of non-transgenic (Non-Tg) ~ 40 days, vehicle-treated (hFUS veh) and niclosamide-treated hFUS mice (hFUS nic) at end stage of disease were subjected to Western blot with anti-S100A4 and anti-MyoG (a); with anti-p-STAT3, anti-p-mTOR, and anti-SQSTM1/p62 (c); and with anti-PDGFR-β and α-SMA (d). GAPDH served as loading control, n = 4 mice/group. Alongside the blots are reported the relative densitometric values. 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)= 12.65 (S100A4), (2, 9) = 8.492 (MyoG), (2, 9) = 12.21 (p-STAT3), (2, 9) = 8.118 (p-mTOR), (2, 9) = 12.04 (SQSTM1/p62), (2, 9) = 10.07 (PDGFR-β), (2, 9) = 14.17 (α-SMA). *p < 0.05 and **p < 0.01 vs. Non-Tg mice or #p < 0.05 and ##p < 0.01 vs. hFUS veh mice. b Real time PCR for MyoG, MyoD, Pax7, and MHC in the gastrocnemius muscles of non-transgenic (Non-Tg) ~ 40 days, vehicle-treated (hFUS veh) and niclosamide-treated hFUS mice (hFUS nic) at end stage of disease. Data are normalized to β-actin and expressed as mean ± S.E.M. of n = 3 mice/group, experiments performed in triplicate. One-way ANOVA with Tukey correction between Non-Tg, hFUS veh, and hFUS nic. F value (DFn, DFd): (2, 6) = 5.790 (MyoG), (2, 6) = 5.648 (MyoD), (2, 6) = 6.155 (Pax7), (2, 6) = 9.657 (MHC). *p < 0.05 vs. Non-Tg mice or #p < 0.05 vs. hFUS veh mice. e Schematic illustration of niclosamide inhibition on multiple signalling pathways, i.e., S100A4, mTOR, SQSTM1/p62, NF-κB, STAT3, α-SMA, N-cadherin, and PDGFRβ dysregulated in ALS patients derived-cells and in ALS mice tissues with overall decrease of inflammatory and fibrotic state