Fig. 3: The α-Syn spreading is reduced in PFF-seeded A53T mice lacking TLR2. | Nature Communications

Fig. 3: The α-Syn spreading is reduced in PFF-seeded A53T mice lacking TLR2.

From: Selective targeting of the TLR2/MyD88/NF-κB pathway reduces α-synuclein spreading in vitro and in vivo

Fig. 3

A53T+/+ mice were crossed with TLR2−/− mice to generate A53TΔTLR2 double transgenic mice. These mice were validated by genetic screening where 324 and 247 bp bands correspond to nTg and A53T Tg mice respectively (marked with arrows). Similarly, 499 and 334 bp bands indicate nTg and TLR2−/− mice respectively (shown with arrows) (a). Spreading of α-syn in SN of PFF-seeded A53T and A53TΔTLR2 mice was compared by assessing total α-syn level in Triton X-100 soluble (b, d) and insoluble (c, e, p = 0.0027 for A53T + PBS vs A53T + PFF and p = 0.014 for A53T + PFF vs A53TΔTLR2+PFF) fractions by immunoblotting. Actin was used as the loading control. Level of pSyn129 in SN (f, g, p = 0.00015 for A53T + PFF vs A53TΔTLR2+PFF) and motor cortex (h, i, p = 0.0003 for A53T + PFF vs A53TΔTLR2+PFF) was monitored by immunostaining (f, h) followed by pSyn129 intensity analysis using Fiji. Two sections from each brain were used for staining and individual values from each section are shown. Two-way ANOVA was performed to determine statistical significance among different groups. *p < 0.05, **p < 0.01, and ***p < 0.001 indicate significance compared to respective groups. Values are given as mean ± SEM (n = 4 animals per group).

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