The motor function of the mice was examined prior to, 24 and 72 h after ischemic onset.

<p><b>A</b>) 24 h after ischemia, all mice demonstrated significant functional deficits that were not affected by sulforaphane treatment. (mean±SD, <i>n</i> = 8–12, t-test or one way ANOVA *<i>p</i>≤0.05, **<i>p</i>≤0.001, ***<i>p</i>≤0.0001; solid black circle vehicle, solid grey circle 5 mg/kg sulforaphane, open box 50 mg/kg sulforaphane). <b>B</b>) 72 h after ischemia, mice treated with sulforaphane were able to complete the round beam task compared to vehicle. All other behavioural outcome measurements had returned to pre stroke baseline levels. (mean±SD, <i>n</i> = 8–12, solid black line vehicle, solid grey line 5 mg/kg sulforaphane, dashed grey line 50 mg/kg sulforaphane, *** F(2,48) = 53.21, <i>p</i>≤0.0001).</p

Infarct volume presented as scattergrams and as their distribution across in anterior and posterior plane at A) 24 h and B) 72 h after ischemia.

<p>Infarct volume and distribution did not change with time. Single or repeated doses of sulforaphane did not protect as infarct volume did not differ in comparison to vehicle treated animals (mean±SD; <i>n</i> = 8–13, one way ANOVA 24 h p = 0.38, 72 h p = 0.93; solid black vehicle, solid gre 5 mg/kg sulforaphane, open box 50 mg/kg sulforaphane). Coronal images of representative cortical infarcts at 0.5 mm from Bregma 72 h after onset in C) vehicle, D) 5 mg/kg sulforaphane and E) 50 mg/kg sulforaphane treated animals.</p

Cell proliferation and glial cell distribution determined by immunostaining 72 h after infarction.

<p>The bar graphs (mean±SD) represent the cell distribution in the peri-infarct, transition and core regions <b>A</b>) the percentage of total cell number per region and <b>B-D</b>) the number of cells/µm², <b>B’</b>) activated microglia (Iba1), <b>C’</b>) proliferating cells (BrdU) and <b>D’</b>) reactive astrocytes (GFAP). No GFAP positive cells were observed in the core region. Sulforaphane treatment only altered the microglial cell number in the peri-infarct region compared to vehicle treatment (F(2,15) = 4.76, <i>p</i>≤0.025) (<i>n</i> = 5–9 in each group, solid black box GFAP astrocytes, solid grey box Iba1 microglia, open box BrdU proliferating cells).</p

Messenger RNA levels of Nrf2 related gene products.

<p><b>A</b>) Nqo1 and Nrf2 (Nfe2l2) expression in brain of naïve mice 24 h after treatment with sulforaphane or vehicle. Nrf2 is significantly increased by 5 mg/kg sulforaphane (mean±SD; <i>n</i> = 3, *<i>p</i>≤0.05; solid black box vehicle, solid grey box 5 mg/kg sulforaphane, open box 50 mg/kg sulforaphane). <b>B</b>) There was no additive effect of sulforaphane on stimulation of the Nrf2 system. Hmox1 and Gsta4 expression was significantly greater in animals 12 h after ischemia and 5 mg/kg sulforaphane treatment. GCLC treatment was increased by ischemia alone. (mean±SD; <i>n</i> = 3, ***<i>p</i>≤0.0001; solid black box sham and vehicle, solid grey box ischemia alone, open box grey diagonal stripes ischemia and 5 mg/kg sulforaphane).</p

Randomisation, blinded outcome assessment, and sample size calculation in systematic reviews of animal studies.

a<p>Summarises the data of six systematic reviews of treatment strategies for acute ischemic stroke. There is an overlap of 18 publications between references <a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.1000245#pmed.1000245-Sena1" target="_blank">[16]</a> and <a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.1000245#pmed.1000245-VanderWorp2" target="_blank">[19]</a>.</p><p>ALS, amyotrophic lateral sclerosis; N/A, data not available; RDS, respiratory distress syndrome.</p

Four types of bias threatening internal validity.

<p>Adapted from <a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.1000245#pmed.1000245-Juni1" target="_blank">[12]</a>,<a href="http://www.plosmedicine.org/article/info:doi/10.1371/journal.pmed.1000245#pmed.1000245-Altman1" target="_blank">[13]</a>.</p