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. 2010 Apr 12;38(15):5130–5140. doi: 10.1093/nar/gkq198

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Published by Oxford University Press (2010).

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 6. — Identification of sFlt1 mRNA proximal intronic cleavage site with the reporter vector pCβS. (A) Schematic representation of the pCβS vector with sFlt1 proximal fragment inserted downstream of the CMV promoter to create pCβs/sFlt1 prox. (B and C) pCβS/sFlt1 prox or pCβs/sFlt1proxmut was transfected into HTR-8/SVNeo cells and then isolated mRNA was probed by RPA with a cRNA derived from the corresponding plasmid. A strong band in (B) lane 1 at ∼170 represents cleavage directed by the putative proximal intronic sFlt1 poly(A) signal sequence. Mutation of the proximal signal leads to loss of the primary cleavage at ∼170 in (C) lane 1. A weak band at ∼190 seen in lane 1 (C), probably represents cleavage directed by a cryptic poly(A) signal sequence. The signal sequence is boxed in sequence above (B and C). Digested RNA samples were run alongside yeast RNA (negative control) and (undigested) probe.