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Fig. 10. Validation of microarray data by immunohistochemistry. Cryosections of
neonatal (P7) cerebellum (A,D,G,J), pre-neoplastic lesions (B,E,H,K) and
tumors (C,F,I,L) from patched+/– mice were stained
with primary antibodies specific for Zic3 (A-C), Pax6 (D-F), Necdin (G-I) or
Hsp105 (J-L) and peroxidase-conjugated secondary antibodies. Staining was
detected using the peroxidase substrate DAB, which yields a dark brown
precipitate. Yellow brackets mark the boundaries of the EGL (panels A,D,G,J),
pre-neoplastic (PN) regions (B,E,H,K) and tumor (C,F,I,L). Expression of all
four proteins is detectable in GCPs within the EGL. Consistent with microarray
data, expression of Zic3 decreases significantly in pre-neoplastic lesions and
is absent from tumors; Pax6 and Necdin are restricted to peripheral regions at
the pre-neoplastic stage and are undetectable in tumors; and Hsp105 expression
increases markedly at the pre-neoplastic stage and decreases somewhat in
tumors.
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