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Figure 1.

3MST activities in six different regions of the rat brain.

3MST activities were measured biochemically in tissue homogenates in the absence (solid bars) and presence of 2-ketobutyric acid (open bars) as described in Methods; N=3.

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Figure 2.

Cellular localization of 3MST using dual-fluorescent immunohistochemical labelling on rat cortical sections.

(a) 3MST immunoreactivity (green) does not colocalize with NeuN-immunoreactive neurons (red). (b) 3MST immunoreactivity colocalizes with GFAP-immunoreactive astrocytes (red). (c) 3MST does not colocalize with OX42-immunoreactive mciroglial cells (red). Scale bar = 50µm. All sections were observed with an Olympus confocal fluorescent microscope.

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Figure 3.

Cellular localization of 3MST using dual-fluorescent immunohistochemical labelling on rat striatal sections.

(a) 3MST immunoreactivity (green) does not colocalize with NeuN-immunoreactive astrocytes (red). (b) 3MST immunoreactivity (green) colocalizes with GFAP-immunoreactive neurons (red). (c) 3MST (green) does not colocalize with OX42-immunoreactive mciroglial cells (red). Scale bar = 50µm. All sections were observed with an Olympus confocal fluorescent microscope. (d) High magnification photomicrograph showing an astrocyte stained positive for both 3MST and GFAP. Scale bar = 10 µm.

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Figure 4.

A TTC-stained sections (2 mm) of a rat forebrain after pMCAO.

The infracted regions are not stained by TTC thus appear white. Photomicrographs presented in Figs. 5 – 7 are taken from the peri-infarct areas at the level of +1mm from Bregma as indicated. a: Cortex; b: corpus callosum; c: striatum.

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Figure 5.

3MST expression in the ipsilateral cortex at various time points after pMCAO.

(A) Representative western blot bands are shown in the top panel. N=4. *p<0.05 against the contralateral side. (B) 3MST immunoreactivity (green) in the ipsilateral cortex 24 h after pMCAO (top row) and sham-operated (bottom row) cortex showing co-localization with GFAP-immunoreactive astrocytes (red). Scale bar = 100µm. (C) 3MST immunoreactivity (green) in the ipsilateral cortex 72 h after pMCAO (top row) and sham-operated (bottom row) cortex showing co-localization with GFAP-immunoreactive astrocytes (red). Scale bar = 100µm.

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Figure 6.

3MST expression in the ipsilateral striatum at various time points after pMCAO.

(A) Representative western blot bands are shown in the top panel. N=4. * p<0.05 against the contralateral side. (B) 3MST immunoreactivity (green) and GFAP-immunoreactive astrocytes (red) in the ipsilateral striatum 24 h after pMCAO (a) and sham-operated striatum (b). Merged photomicrographs (left) show co-localization of 3MST and GFAP immunoreactivity (yellow). Scale bar = 100µm. (C) 3MST immunoreactivity (green) and GFAP-immunoreactive astrocytes (red) in the ipsilateral striatum 72 h after pMCAO (a) and sham-operated striatum (b). Merged photomicrographs (left) show co-localization of 3MST and GFAP immunoreactivity (yellow). Scale bar = 100µm.

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Figure 7.

3MST expression in the corpus callosum.

(A) 3MST immunoreactivity (green) and GFAP-immunoreactive astrocytes (red) in the ipsilateral corpus callosum 24 h after pMCAO (a) and sham-operated corpus callosum (b). Merged photomicrographs (left) show co-localization of 3MST and GFAP immunoreactivity (yellow). Scale bar = 100µm. (B) 3MST immunoreactivity (green) and GFAP-immunoreactive astrocytes (red) in the ipsilateral corpus callosum 72 h after pMCAO (a) and sham-operated corpus callosum (b). Merged photomicrographs (left) show co-localization of 3MST and GFAP immunoreactivity (yellow). Scale bar = 100µm.

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Figure 7 Expand