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Fig. 8 | Journal of Neuroinflammation

Fig. 8

From: RNS60 exerts therapeutic effects in the SOD1 ALS mouse model through protective glia and peripheral nerve rescue

Fig. 8

RNS60 activates the p-Akt pro-survival pathway in the LSC and sciatic nerves of C57BL/6-SOD1G93A mice. a, b Representative immunoblots performed on the ventral portion of LSC of NTG mice or transgenic C57BL/6-SOD1G93A mice treated with NS or RNS60, at 20 weeks of age, and relative quantification. There was an increase of Akt phosphorylation (at Ser473) in RNS60 treated mice compared to NS treated mice. This was paralleled by increased phosphorylation of GSK3β at Ser9 (c, d), one of the downstream targets of activated Akt. e–g Representative images of the co-localization of p-Akt and GFAP immunostaining. P-Akt (red) was highly expressed specifically in the MNs of NTG mice (e). In SOD1G93A mice treated with NS, the number of p-Akt labeled MNs decreased while few of the highly reactive astrocytes (green) co-localized with p-Akt (f). Colocalization of Akt and GFAP was exacerbated in the SOD1G93A mice treated with RNS60 (G, arrows). The inset in G show enlarged reactive astrocytes expressing p-Akt and showing a spheroid or less ramified morphology (n = 3 animals per group). h–j Representative images of the co-localization of p-Akt (red) and CD68 (green) immunostaining. In SOD1G93A mice treated with RNS60, few CD68-positive microglial cells colocalized with p-Akt (arrow), while this phenomenon was not detected in SOD1G93A mice treated with vehicle (i). The inset in J show enlarged macrophagic-microglial cells expressing p-Akt (n = 3 animals per group). k, l Decreased levels of Ser473 Akt phosphorylation were found in the sciatic nerves of SOD1G93A mice treated with NS but not in RNS60 treated mice compared to NTG. Bar graphs represents mean ± SEM as a percentage versus NTG controls (n = 5 animals per group). All data were statistically analyzed using one-way ANOVA followed by post hoc Fisher’s LSD. *p < 0.05, **p < 0.01, ***p < 0.001, n.s. = not significant

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