Immunohistochemistry was performed principally by the following methods previously described27 (link). Sections were pre-treated with Proteinase K before TUNEL staining using ApopTag peroxidase in situ apoptosis detection kit (Millipore). The staining was amplified using biotin-conjugated secondary antibodies, VECTASTAIN ABC system (Vector) and TSA Plus system (Perkin Elmer). Monoclonal anti-BrdU (1:100, Clone 3D4, BD Biosciences, 555627), anti-Calbindin (1:1000, Abcam, ab82812), anti-Tuj1 (1:1,000, Abcam, ab78078), anti-Nestin (1:50, rat-401, DSHB), anti-FLAG M2 (1:50, Sigma, F1804) and anti-NeuN (1:500, Clone A60, Millipore, MAB377) antibodies were used. Polyclonal anti-HSF1X50 (link) (1:500), anti-HSF1 antibody (1:200, C-19, SCBT, sc-8061) and anti-Cutl1 (1:100, M-222, SCBT, sc-13024) antibodies were also used. The HSF1 expression pattern in Supplementary Fig. 5 was consistently obtained by using both HSF1X and HSF1 C-19 antibodies.
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