Fig 1.
Angle transformation for laterality correction.
Cells in the gray matter between 90° and 270° (left side; white cells) were mirrored onto the right side of the gray matter. The angle of cells originally on the right side (black cells) was not altered.
Fig 2.
Representative confocal microscopy images of immunostained spinal cord cross-sections from each genotype.
(A) Full mosaic-merge images. Images were adjusted for brightness and contrast for illustrative purposes only. (B) Corresponding insets from A to show examples of positively stained cells more clearly. Scale bars = 500 μm (A) or 200 μm (B).
Fig 3.
Number of ChAT+ (A-C), GAD-67+ (D), Calbindin+ (E), Parvalbumin+ (F), Parvalbumin+−Calbindin+ (G), Parvalbumin+−GAD-67+ (H), and ChAT+−GAD-67+ (I) cells per biological replicate. The number of cells analysed is the sum of immunostained positive cells from eight spinal cord cross-sections (two sections per lumbar level L3–L6) per biological replicate. Data are shown as mean ± standard deviation and were analysed by an ordinary one-way ANOVA. N = 3 biological replicates per genotype.
Fig 4.
(A-I) Normalised distances of cells relative to the central canal.
The distances were normalised to the size of the gray matter to account for size differences between spinal cord cross-sections. The normalised distance is a unitless fraction between 0 (closest to the central canal) and 1 (farthest from the central canal). Data are from eight spinal cord cross-sections (two sections per lumbar level L3–L6) per biological replicate. Boxes represent the interquartile range with a line in the middle representing the median. The whiskers represent the minimum and maximum values. Data were analysed by a nested one-way ANOVA. N = 3 biological replicates per genotype.
Fig 5.
(A-D) Spatial distribution of immunopositive cells displayed as rose diagrams (i.e., circular histograms) of cell angles relative to the central canal. All left side angles were mirrored to the right side to avoid displaying artificial laterality due to the inability to distinguish the true left and right sides of free-floating spinal cord cross-sections. The length of each bar represents the percentage of cells found at a particular angle (not the distance from the central canal; distance information is ignored in these plots). Data are from eight spinal cord cross-sections (two sections per lumbar level L3–L6) per biological replicate. N = 3 biological replicates per genotype.
Fig 6.
(A-C) Spatial distribution of double-positive cells displayed as rose diagrams (i.e., circular histograms) of cell angles relative to the central canal. All left side angles were mirrored to the right side to avoid displaying artificial laterality due to the inability to distinguish the true left and right sides of free-floating spinal cord cross-sections. The length of each bar represents the percentage of cells found at a particular angle (not the distance from the central canal; distance information is ignored in these plots). Data are from eight spinal cord cross-sections (two sections per lumbar level L3–L6) per biological replicate. N = 3 biological replicates per genotype.
Fig 7.
(A-E) Summary of post hoc pairwise comparisons for cell types that displayed significant main effects. The left column shows heatmaps of false discovery rate (FDR) values to indicate statistical significance. The right column shows the mean angle of the cells (circles) and the 95% confidence interval (error bars) for each genotype.
Table 1.
Circular means and variances from cell angle data for cell types displaying statistically significant differences between genotypes.