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Figure 2 | BMC Molecular Biology

Figure 2

From: A family of splice variants of CstF-64 expressed in vertebrate nervous systems

Figure 2

The βCstF-64 protein is expressed predominantly in mouse brain. A) Whole cell extracts from HeLa cells transfected with 800 ng of either 3XFLAG (lane 1), 3XFLAG-CstF-64 (lane 2) or 3XFLAG-βCstF-64 (lane 3) were subjected to immunoblot analysis using anti-βCstF-64 (upper panel), anti-FLAG (middle panel), or anti-α-Tubulin (lower panel) antibodies. The apparent molecular weights of 3XFLAG-CstF-64, 3XFLAG-βCstF-64, and tubulin are indicated at left. B) Protein immunoblot detection of βCstF-64 in various mouse tissues. Nuclear extracts from indicated mouse tissues were subjected to immunoblot analysis using pre-immune serum (lane 8), peptide-blocked anti-βCstF-64 antibody (lane 9), anti-βCstF-64 antibody (upper panel, lanes 1–7), or anti-CstF-64 antibody (lower panel, lanes 1–7). The apparent molecular weights of CstF-64 and βCstF-64 are indicated at left. C) 2D-PAGE analysis of mouse brain and liver nuclear extracts. Nuclear extracts were resolved in the first dimension by isoelectric focusing on a pH 3–10 immobiline strip as indicated. The proteins were then resolved in the second dimension by denaturing PAGE, transferred to nitrocellulose membrane and probed with the anti-CstF-64 antibody (panels a and c) or anti-βCstF-64 antibody (panels b and d). The arrows denote CstF-64 and βCstF-64. The asterisk in panels a and c indicates a pattern that may be αCstF-64.

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