Characterization of TDP-43 LLPS-deficient knock-in mice. (A) Amino acid conservation score along an alignment of NTD and RRM1/2 of TDP-43 from 50 eukaryotic species. The histogram shows conservation scores for each position of the alignment; the red line shows the 5-aa moving average. (B) Representative images of WT, TDPΔCR^+/−, and TDPΔCR^+/+ mice at P0. (C–I) Quantification of the indicated organ weight of WT, TDPΔCR^+/−, and TDPΔCR^+/+ mice (n = 4–9 mice per group) at P0. (J) Representative hematoxylin and eosin staining of lung tissues from WT and TDPΔCR^+/+ mice at P0. (K) PCR and DNA-seq analysis of WT and mutant mouse TDP-43 alleles in WT, hemizygous (TDPΔCR^+/−), or homozygous (TDPΔCR^+/+) TDPΔCR mice. (L) Forelimb and hindlimb grip strength of WT and TDPΔCR^+/− mice (n = 5–14 mice per group) at 3 mo old or 10 mo old. (M) Rotarod performance of WT mice (n = 13) and TDPΔCR^+/− (n = 10 mice). (N–P) The distance traveled (N), velocity (O), and time spent in the center of apparatus (P) in the open field test. WT mice (n = 13) and TDPΔCR^+/− (n = 17 mice). (Q) Time spent in the open arm in the elevated plus maze. WT mice (n = 9) and TDPΔCR^+/− mice (n = 10). (R) Time spent with each object and preference ratio for an unfamiliar object in the two different sessions of the novel object recognition test. WT mice (n = 6) and TDPΔCR^+/− mice (n = 6). (S) Freezing behavior of WT mice (n = 8) and TDPΔCR^+/− mice (n = 9) in contextual fear learning and cue-dependent fear learning during the fear-conditioning test. CS, conditioned stimulus. Data are mean ± SEM; one-way ANOVA followed by Tukey’s multiple comparisons test (C–I), two-tailed Student’s t test (L–S).