Fig. 4: Assessment of non-motor and motor symptoms in mice intranasally treated with O-αSyn.

A–C Behavioral data from open field test. Average speed (A), time spent in the center zone (B) and number of entries into the center zone (C) were auto-measured by ANY-maze software. D, E Behavioral data from elevated plus maze test. Time spent in the open arms (D) and number of entries into the open arms (E) were obtained from each 5-minute trail. F Latency to find the food pellets in buried food test. G Data from novel object recognition test. Recognition index were defined as the ratio of time spent exploring the novel object to the total time spent exploring both objects. H Immobility time of the mice in tail suspension test. I Withdrawal threshold measured in von Frey test. J Latency of tail-flick behavior in tail flick test. K–M Locomotor behavior of the mice in treadmill test. Representative traces (K) of the mice treated with PBS or O-αSyn on the treadmill were showed, and time spent (L) and distance traveled (M) in the front zone were further analyzed using ANY-maze software. N Average speed in the open field test was analyzed automatically using ANY-maze software. O Latency to fall off the apparatus in Rotarod test. P The descending time in pole test. Q Quantification of TH⁺ cells in the substantia nigra of the mice treated with PBS or O-αSyn. Unpaired two-tailed Student’s t test, n = 18 for (A–J); n = 6 for (K–M): Two-way repeated measures ANOVA, n = 18 for 1-month data in (N–P); n = 12 for 6-month data in (N–P); n = 8 for 15-month data in (N–P); Unpaired two-tailed Student’s t test, n = 4 for Q, ^*p < 0.05, ^**p < 0.01, ^***p < 0.001.