Figure 5
From: Ebselen analogues delay disease onset and its course in fALS by on-target SOD-1 engagement
MR6-26-2 decreased misfolded SOD1 species and delayed the disease onset of SOD1G93A mice. (A,B) Onset (A) and survival (B) curves of the control or MR6-26-2 treated female SOD1G93A mice plotted over time (n = 20, each). The mean ages for onset or survival are shown with SD. (C) Changes in clasping score of the control or MR6-26-2 treated SOD1G93A mice plotted over time (n = 20, each). The data were analyzed by two-way ANOVA following posthoc multiple comparisons with Šidák correction. (D,E) Decrease of insoluble SOD1 species in the spinal cords of MR6-26-2 treated SOD1G93A mice at 120 days old. A representative immunoblotting image is shown (D), and the relative expressions of insoluble SOD1 per soluble SOD1 were quantified and plotted as mean ± SEM (E). (F,G) SOD1 oligomers produced by aberrant disulfide bonds (hSOD1 S-S oligomers) were decreased in the spinal cords of MR6-26-2 treated SOD1G93A mice at 120 days old. A representative immunoblotting image indicates the hSOD1 S-S oligomers (a bold line) (F). Relative expressions of hSOD1 S-S oligomers were quantified and plotted as mean ± SEM (G). (H–J) Immunofluorescent analyses of lumbar spinal cord sections of early symptomatic SOD1G93A mice showed the decrease of misfolded SOD1 species, detected using a C4F6 antibody that specifically recognizes misfolded SOD1, by the MR6-26-2 treatment. Motor neurons were identified using an anti-choline acetyltransferase (ChAT) antibody. Representative immunofluorescent image is shown (H), and the relative fluorescence intensity of C4F6 (I) or the number of ChAT-positive motor neurons (J) in the ventral horn averaged per mouse was quantified and plotted as mean ± SD. Each dot represents the section used for the quantification (I,J). Scale bar: 50 µm.
