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. Author manuscript; available in PMC: 2022 Jun 6.
Published in final edited form as: Nature. 2022 Feb 14;602(7898):647–653. doi: 10.1038/s41586-022-04396-8

Extended Data Fig. 10. Fluorescence imaging and extended behavior test results of clemastine fumarate and miconazole-treated mice, and preliminary behavior tests performed on both sexes, used to determine continued studies on males for the study.

Extended Data Fig. 10

a-i, 4EP+/− mice were administered vehicle (control), clemastine fumarate (clem.), or miconazole and then behavior tested and imaged.. a, Representative images of CC1 (green), NG2 (red), and OLIG2 (blue) staining in 4EP+/− mice with or without treatment. Representative of group of individual mice used for quantification: 4EP- control=7, treated=10; 4EP+ control=6, treated=6, which came from two cohorts (Scale bar 100µm). b, Quantification of fluorescent imaging, presented as a ratio of CC1+/NG2+ cells (from left to right n=7, 10, 6, 6, 4, 4). The first four columns are also shown in Main Fig. 4i (Cont 4EP-vs 4EP+ p=0.04; 4EP+Cont vs 4EP+Clem p=0.01). c-d, Confirmation that treatment does not reduce 4EPS levels with clemastine (c) (Control. 4EP- n=4; Control. 4EP+ n=6, Clem 4EP- n=3; Clem 4EP+ n=4) or miconazole (d) (Control. 4EP- n=4; Control. 4EP+ n=4, Mic 4EP- n=4; Mic 4EP+ n=5) treatment. e, Extended EPM results (Control 4EP- n=13; Control 4EP+ n=13; Clem 4EP- n=12; Clem 4EP+ n=14). f, Extended open field results (Control 4EP- n=15; Control 4EP+ n=17; Clem 4EP- n=17; Clem 4EP+ n=17). g-i, Behavioral results for mice treated with miconazole, including g, Open field (Control 4EP- n=24; Control 4EP+ n=25; Mic. 4EP- n=18; Mic. 4EP+ n=20)(left graph: 4EP-Cont vs 4EP+ Cont p=0.02; 4EP+cont vs 4EP+Mic p=0.008)(middle graph: 4EP-Cont vs 4EP+ Cont p=0.02; 4EP+cont vs 4EP+Mic p=0.005). h, EPM (Control 4EP- n=24; Control 4EP+ n=25; Mic. 4EP- n=15; Mic. 4EP+ n=17)(left graph: 4EP-Cont vs 4EP+ Cont p=0.002; 4EP+cont vs 4EP+Mic p=0.03)(right graph: 4EP-Cont vs 4EP+Cont p=0.0009). i, Marble Burying (Control 4EP- n=24; Control 4EP+ n=26; Mic. 4EP- n=21; Mic. 4EP+ n=21)(p=0.001) j, Left panel: EPM, time spent in open arms over time spent in closed arms. Right panel: time spent at terminus (outer 1/3 of open arms) (Males 4EP- n=17; Males 4EP+ n=17; Females 4EP- n=20; Females 4EP+ n=21)p=0.02. k, Open field test, time spent in center area over time spent in thigmotaxis left), time spent in center (right) (Males 4EP- n=16; Males 4EP+ n=14; Females 4EP- n=18; Females 4EP+ n=13)p-values left to right: 0.005, 0.003. l, Number of marbles buried in marble burying test (Males 4EP- n=24; Males 4EP+ n=23; Females 4EP- n=17; Females 4EP+ n=26)p=0.03. Abbreviations: NG2, neural/glial antigen 2; CC1, antibody (anti-adenomatous polyposis coli (APC) clone) that binds mature oligodendrocyte marker; OLIG2, oligodendrocyte transcription factor 2; clem, clemastine fumarate; mic, miconazole. Two independent cohorts of mice from multiple litters were used for each experiment in this figure. Data represent mean ± SEM analyzed by two-way ANOVA with Dunnett multiple comparison to 4EP+ group (panels b-i) or Sidak multiple comparison between 4EP+/− groups within each sex (panels j-l). * p ≤ 0.05, ** p ≤ 0.01.