(A) P62 (green), α-syn (red), and TH (blue) co-staining in the midbrain sections of 18-month-old nTg, as well as 1- and 18-month-old A53T mice. Arrowheads point to areas with substantial overlap of P62 and α-syn staining. Scale bar: 10 μm
(B) The quantification of P62 staining intensity in the TH-positive neurons of 1-month-old nTg and A53T mice (n = 3 animals per genotype; n ≥ 20 neurons per animal). Data were presented as Mean ± SEM. ***p<0.001
(C) Western blot analysis shows the expression of P62 in the midbrain homogenate of 1-month-old A53T and littermate nTg mice. TH was used as loading control. Bar graph depicts the P62 expression levels normalized with TH in the midbrain homogenate of 1-month-old A53T and littermate nTg mice (n = 3 per genotype). Data were presented as Mean ± SEM. *p<0.05
(D) LC3 (green), human α-syn (red), and TH (blue) co-staining in the midbrain section of 1- month-old nTg and littermate A53T mice. Scale bars: 50 μm (low magnification images) and 10 μm (high magnification images).
(E) The size of LC3-positive puncta in the TH-positive neurons of 1-month-old nTg and A53T mice (n = 3 animals per genotype; n ≥ 20 neurons per animal). Data were presented as Mean ± SEM. *p<0.05
(F) Western blot shows the expression of LC3BI and LC3BII in the midbrain homogenate of 1- month-old A53T and nTg mice.
(G) LAMP2 (green), α-syn (red), and TH (blue) co-staining in the midbrain sections of 1-month- old nTg and littermate A53T mice. Scale bar: 10 μm
(H) The density of LAMP2-positive puncta in the TH-positive neurons of 1-month-old nTg and A53T mice (n = 3 animals per genotype; n ≥ 20 neurons per animal). Data were presented as Mean ± SEM. ***p<0.001
(I) Western blot analysis shows the expression of LAMP2 in the midbrain homogenate of 1-month-old A53T and littermate nTg mice. TH was used as loading control. Bar graph depicts the LAMP2 expression levels normalized with TH in the midbrain homogenate of 1-month-old A53T and littermate nTg mice (n = 3 per genotype). Data were presented as Mean ± SEM. *p<0.05