Phosphorylated α-synuclein lesions in the α-Gal A-deficient pons co-localize with ubiquitin and LC3. Sagittal brain sections from male 20- to 24-month-old α-Gal A +/0 (a-d) or -/0 (b-l) mice were immunolabeled with antibodies against phosphorylated α-synuclein (a, e, i) and ubiquitin (b, f) or LC3 (j). Colored arrowheads indicate large lesions containing phosphorylated α-synuclein, ubiquitin, or LC3, and arrows indicate co-localization of large phosphorylated α-synuclein-containing lesions with either ubiquitin (h) or LC3 (l). Co-localization analysis was performed using the thresholded Manders test. Manders co-localization between ubiquitin (M1) and phosphorylated-α-synuclein (M2) was calculated to be (tM1 = 0.506 and tM2 = 0.856, Costes p-value = 1, n = 3), indicating strong co-localization of phosphorylated-α-synuclein signal with ubiquitin in the pons of α-Gal A -/0 mice. Manders co-localization between phosphorylated-α-synuclein (M1) and LC3 (M2) was calculated to be (tM1 = 0.499 and tM2 = 0.765, Costes p-value = 0.958, n = 3), indicating strong co-localization of LC3 signal with phosphorylated-α-synuclein in the pons of α-Gal A -/0 mice. Scale bars = 50 microns.