Extended Data Fig. 10: Knockdown of Kcnn2 in layer II/III neurons in M1 improves motor learning deficits in PAE mice.

a, Timeline of the experiment. b, Representative image of a brain that received Kcnn2 (or control) knockdown (visualized by co-expressed GFP) in the motor area (outlined by red rectangles). c, Immunohistochemistry for Kcnn2 (red) shows that Kcnn2 shRNA, but not control shRNA, suppresses the increase in Kcnn2 expression in layer III neurons in M1 in PAE mice. Arrowheads indicate Kcnn2⁺ cells among GFP⁺ electroporated cells. d, Percentage of Kcnn2⁺ cells among GFP⁺ electroporated cells in the indicated experimental groups. **P = 0.001 by two-tailed Student’s t-test (n = 10 per group). (e, f) Motor learning deficits in PAE mice, revealed by lower success rate (e) and learning index (f) in the single pellet reaching test, are mitigated by Kcnn2 knockdown in layer II/III neurons in M1. A significant interaction was observed between the effects of condition (treatment plus shRNA) and trial (e); F(1,11) = 2.80, P = 0.01 by two-way repeated measures ANOVA, *P < 0.05, **P < 0.01 by simple main effect test [PAE (Kcnn2 shRNA⁻) vs PAE (Kcnn2 shRNA⁺)], and between treatment and shRNA (f); F(1,24) = 5.55, P = 0.03 by two-way ANOVA, **P < 0.05, ***P < 0.005 by simple main effect test (n = 10 animals per group). Graph shows mean ± SEM. In box plots (d, f), the line within the box indicates the median, and the upper and lower edges of the box represent the 25th and 75th percentiles, respectively. The upper and lower whisker boundaries indicate the 10th and 90th percentiles, respectively, and dots indicate outliers.