Figure S4.
MAZ51 improves early post-stroke motor outcomes. (A) Experimental timeline of behavioral analysis in MAZ51-treated tMCAO mice (i.p. injected on days 0, 2, 4, and 6). Mice were tested in rotarod, open field, and foot fault for post-stroke motor function across days 1, 3, 5, and 7 (created using https://BioRender.com). (B) The ladder rung test was used to measure the total number of foot faults to assess locomotor ability between control-treated tMCAO mice and MAZ51-treated tMCAO mice at days 1, 3, 5, and 7 (n = 6 mice per group; mean ± SEM, *P < 0.05, two-way ANOVA with repeated measures and Sidak’s multiple comparison test). (C) An open-field behavioral test was used to track the ability of the mouse to move to the peripheral wall after initial placement. Healthy/sham mice within seconds move to the peripheral wall. tMCAO mice have difficulty moving in a straight line due to single-side paralysis/circling behavior and have elevated time to reach the periphery. (D) Latency to reach the periphery of the open-field box was measured between all groups, including control-treated tMCAO mice and MAZ51-treated tMCAO mice, at days 1, 3, 5, and 7 (n = 6 mice per group; mean ± SEM, **P ≤ 0.05, two-way ANOVA with repeated measures and Sidak’s multiple comparison test). (E) Representative track maps between tMCAO with control or MAZ51 at days 1 and 7 for comparison. The blue box indicates the center of the box and the red line indicates the peripheral boundary of the box. While control-treated tMCAO mice showed difficulties in movement, MAZ51-treated tMCAO mice showed improved locomotor activities at day 1. (F) Latency duration on the rotarod was measured between control-treated tMCAO mice and MAZ51-treated tMCAO mice at days 1, 3, 5, and 7 (n = 6 mice per group; mean ± SEM, *P < 0.05, two-way ANOVA with repeated measures and Sidak’s multiple comparison test). (G) Timeline summary of motor function effects of MAZ51 treatment in relation to relative peak CP lymphangiogenesis (day 7). Early improvements in motor recovery (days 1–3), but late deficit in rotarod (day 7) could be associated with sustained MAZ51 inhibition of lymphangiogenesis. Peak post-stroke lymphangiogenesis typically occurs at the CP on day 7 after stroke (created using https://BioRender.com). Source data are available for this figure: SourceData FS4.

MAZ51 improves early post-stroke motor outcomes. (A) Experimental timeline of behavioral analysis in MAZ51-treated tMCAO mice (i.p. injected on days 0, 2, 4, and 6). Mice were tested in rotarod, open field, and foot fault for post-stroke motor function across days 1, 3, 5, and 7 (created using https://BioRender.com). (B) The ladder rung test was used to measure the total number of foot faults to assess locomotor ability between control-treated tMCAO mice and MAZ51-treated tMCAO mice at days 1, 3, 5, and 7 (n = 6 mice per group; mean ± SEM, *P < 0.05, two-way ANOVA with repeated measures and Sidak’s multiple comparison test). (C) An open-field behavioral test was used to track the ability of the mouse to move to the peripheral wall after initial placement. Healthy/sham mice within seconds move to the peripheral wall. tMCAO mice have difficulty moving in a straight line due to single-side paralysis/circling behavior and have elevated time to reach the periphery. (D) Latency to reach the periphery of the open-field box was measured between all groups, including control-treated tMCAO mice and MAZ51-treated tMCAO mice, at days 1, 3, 5, and 7 (n = 6 mice per group; mean ± SEM, **P ≤ 0.05, two-way ANOVA with repeated measures and Sidak’s multiple comparison test). (E) Representative track maps between tMCAO with control or MAZ51 at days 1 and 7 for comparison. The blue box indicates the center of the box and the red line indicates the peripheral boundary of the box. While control-treated tMCAO mice showed difficulties in movement, MAZ51-treated tMCAO mice showed improved locomotor activities at day 1. (F) Latency duration on the rotarod was measured between control-treated tMCAO mice and MAZ51-treated tMCAO mice at days 1, 3, 5, and 7 (n = 6 mice per group; mean ± SEM, *P < 0.05, two-way ANOVA with repeated measures and Sidak’s multiple comparison test). (G) Timeline summary of motor function effects of MAZ51 treatment in relation to relative peak CP lymphangiogenesis (day 7). Early improvements in motor recovery (days 1–3), but late deficit in rotarod (day 7) could be associated with sustained MAZ51 inhibition of lymphangiogenesis. Peak post-stroke lymphangiogenesis typically occurs at the CP on day 7 after stroke (created using https://BioRender.com). Source data are available for this figure: SourceData FS4.

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