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Supplementary Figures
Figure S1: SLC25A12, AUTS2 and PRKCB1 mRNA levels in the human
brain during early development.
Sagittal sections of 8-week-old human fetal brain hybridized with SLC25A12 (a1
and a3), AUTS2 (b1 and b3) and PRKCB1 (c1 and c3) antisense radioactive
riboprobes or stained with toluidine blue (a2, a4, b2, b4, c2 and c4).
(a1, b1 and c1) SLC25A12 transcripts (a1) were more numerous in the
telencephalon (tel) than in the cerebellum anlagen (Cb), whereas AUTS2 (b1)
and PRKCB1 (c1) transcripts were evenly distributed in these two regions.
(a2, b2 and c2) Telencephalic enlargement of (a1, b1 and c1) showing that the
decreasing gradient of expression from posterior to anterior observed for
SLC25A12 (a2) was not observed for either AUTS2 (b2) or PRKCB1 (c2).
ge, ganglionic eminence; mo, medulla oblongata and oe, olfactory epithelium.
Scale bars (a1, a2, b1, b2, c1, c2) indicate 1 mm and scale bars (a3, a4, b3, b4,
c3, c4) indicate 0.25 mm.
Figure S2: Normalized expression of SLC25A12, AUTS2 and PRKCB1 in 8W
human embryo brain.
SLC25A12, AUTS2 and PRKCB1 mRNA levels were compared, taking absolute
radioactivity levels into account. The intensity scale is indicated in color.
Scale bars indicate 1 mm.
Figure S3: SLC25A12, PRKCB1 and AUTS2 expression in temporal cortex
at mid gestation
Coronal sections of 23-week-old human fetal brains hybridized with SLC25A12
antisense radioactive riboprobe (b2 and c2), with PRKCB1 antisense radioactive
riboprobe (b3 and c3) and with AUTS2 antisense radioactive riboprobe (b4 and
c4), or stained with toluidine blue (b1 and c1).
(a) Schematic diagram of the fetal brain sections used, with sulci present at 23W.
(b)
Identification
of
molecular
gradients
in
the
fusiform
gyrus
and
parahippocampal gyrus at 23W. Section stained with toluidine blue (b1).
Expression of SLC25A12 (b2), PRKCB1 (b3) and AUTS2 (b4) in the temporal
cortex.
(c) Enlargements of hippocampus as illustrated in (b) for SLC25A12 (c2),
PRKCB1 (c3) and AUTS2 (c4).
Arrows in (b1) indicate sulci as in the schematic diagram (a).
dentate gyrus (dg), ganglionic eminences (ge), hippocampus (hipp), thalamus
(th), parahippocampal gyrus (para hipp).
Scale bars indicate 1 mm.
Figure S4: SLC25A12 mRNA levels in human cerebellum at mid-gestation
(21W).
Coronal sections of human 21W fetal cerebellum hybridized with SLC25A12
antisense radioactive riboprobe (a) or stained with toluidine blue (b).
SLC25A12 transcripts are present in the cerebrocerebellum (cc), in vestibular
nuclei (vn) and in deep cerebellar nuclei (dcn), but not in the pons.
Scale bar indicates 1 mm.
Figure S5: Slc25a12 mRNA levels in mice.
Coronal sections of embryonic day 14 (E14) mouse brain (a) and postnatal day
21 (P21) mouse brain (b) hybridized with Slc25a12 antisense radioactive
riboprobe.
(a) Slc25a12 transcripts are present in the cortical plate (cp), ganglionic
eminence (ge) and retina (r).
(b) Slc25a12 transcripts are present in the cortex (co) and hippocampus (hipp)
but not in the internal capsule (ic).
Scale bar (a) indicates 0.5 mm and scale bar (b) indicates 1 mm.
Figure S6: shRNA-Slc25a12 efficacy in a mouse neuroblastoma cell line
(N18).
(a-b) Confocal slices of N18 transfected with Slc25a12-EGFP vector (a) or
cotransfected with Slc25a12-EGFP and shRNA-Slc25a12 constructs (b). Scale
bar indicates 15 m.
(c) Slc25a12 mRNA level was quantified by quantitative real-time RT-PCR (c1)
and Slc25a12-EGFP fluorescent intensity was quantify with LSM Image Browser
software (c2). Note that both Slc25a12 transcript level and Slc25a12-EGFP
fluorescence are downregulated in silenced N18 by shRNA-Slc25a12 vector (**;
p<0.001).