Figure S3.
Pax5 mutant mice display poor motor coordination and ASD-related behavioral abnormalities. All behavioral experiments (A–K) were performed with Pax5+/+, Pax5+/−, and Pax5R31Q/− mice (n ≥ 9 per genotype) at the age of 2–4 mo. (A) ErasmusLadder. The time required for short or long steps is shown as mean value with SEM. (B) LocoMouse. Left: The illustration indicates the measured parameters. Top right: Stride length for each individual paw. Bottom left: The body axis swing reflects the angular difference of the central body axis between the placement of the right-front (RF) and left-hind (LH) diagonal paws and the left-front (LF) and right-hind (RH) diagonal paws. Bottom middle-right: The distances between the front paws (middle) or hind paws (right) during locomotion, as indicated by the dotted line in the cartoon. (C–E) Three-chamber sociability and social novelty test. (C) Illustration of the setup with a single “stranger 1” mouse in one cup (sociability test) or with two stranger mice (“stranger 1 and 2”) in separate cups (social novelty test). (D) Sociability test. Left: Total time spent in the perimeter of the cup with stranger 1 mouse or the empty cup (object). Right: Mean length of bouts actively exploring stranger 1 mouse. (E) Social novelty test. Left: Total time spent in the perimeter of the cup with stranger 1 or stranger 2. Right: Mean length of bouts actively exploring stranger mouse 1 and 2. (F) Marble burying test. Left: Total time spent burying. Right: Mean length of burying bouts. (G) Self-grooming behavior. Left: Total time spent grooming. Right: Mean time of short and long grooming bouts, respectively. Short bouts were defined as 0–3.5 s, long bouts as >3.5 s. (H) Open field test. Total time spent at the periphery or in the center of the arena. (I) Elevated plus maze. Left: The illustration indicates the closed and open arms of the platform. Right: Time spent in the open and closed arms as well as in the center. (J and K) Eyeblink conditioning. (J) Examples of individual traces during US-only (top) and CS-US (bottom) trials (see Video 3 for explanation of the trial types). (K) Quantification of valid and invalid trials during US-only and CS-US–only trials (see Materials and methods for detailed description of the experiment). The data (B and D–I) are shown as mean values with SEM (n ≥ 12 per genotype). Two-way repeated-measures ANOVA with Dunnett’s multiple comparisons test (A), ANOVA with Dunnett’s or Dunnett’s T3 multiple comparisons test (B, F, and G), two-way ANOVA with Tukey’s multiple comparisons test (D, E, H, and I); *, P < 0.05; **, P < 0.01; ***, P < 0.001. For detailed statistical information (B and D–I), see Table S6. Each dot represents one mouse.

Pax5 mutant mice display poor motor coordination and ASD-related behavioral abnormalities. All behavioral experiments (A–K) were performed with Pax5+/+, Pax5+/−, and Pax5R31Q/− mice (n ≥ 9 per genotype) at the age of 2–4 mo. (A) ErasmusLadder. The time required for short or long steps is shown as mean value with SEM. (B) LocoMouse. Left: The illustration indicates the measured parameters. Top right: Stride length for each individual paw. Bottom left: The body axis swing reflects the angular difference of the central body axis between the placement of the right-front (RF) and left-hind (LH) diagonal paws and the left-front (LF) and right-hind (RH) diagonal paws. Bottom middle-right: The distances between the front paws (middle) or hind paws (right) during locomotion, as indicated by the dotted line in the cartoon. (C–E) Three-chamber sociability and social novelty test. (C) Illustration of the setup with a single “stranger 1” mouse in one cup (sociability test) or with two stranger mice (“stranger 1 and 2”) in separate cups (social novelty test). (D) Sociability test. Left: Total time spent in the perimeter of the cup with stranger 1 mouse or the empty cup (object). Right: Mean length of bouts actively exploring stranger 1 mouse. (E) Social novelty test. Left: Total time spent in the perimeter of the cup with stranger 1 or stranger 2. Right: Mean length of bouts actively exploring stranger mouse 1 and 2. (F) Marble burying test. Left: Total time spent burying. Right: Mean length of burying bouts. (G) Self-grooming behavior. Left: Total time spent grooming. Right: Mean time of short and long grooming bouts, respectively. Short bouts were defined as 0–3.5 s, long bouts as >3.5 s. (H) Open field test. Total time spent at the periphery or in the center of the arena. (I) Elevated plus maze. Left: The illustration indicates the closed and open arms of the platform. Right: Time spent in the open and closed arms as well as in the center. (J and K) Eyeblink conditioning. (J) Examples of individual traces during US-only (top) and CS-US (bottom) trials (see Video 3 for explanation of the trial types). (K) Quantification of valid and invalid trials during US-only and CS-US–only trials (see Materials and methods for detailed description of the experiment). The data (B and D–I) are shown as mean values with SEM (n ≥ 12 per genotype). Two-way repeated-measures ANOVA with Dunnett’s multiple comparisons test (A), ANOVA with Dunnett’s or Dunnett’s T3 multiple comparisons test (B, F, and G), two-way ANOVA with Tukey’s multiple comparisons test (D, E, H, and I); *, P < 0.05; **, P < 0.01; ***, P < 0.001. For detailed statistical information (B and D–I), see Table S6. Each dot represents one mouse.

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