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. 2006 May 26;2(5):e48. doi: 10.1371/journal.ppat.0020048

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© 2006 Bracha et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited.

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(A) Diagrams of the Ehlgl1-containing plasmids pB33 and pAY. The 473 bp Ehap-a promoter, SINE and T-rich (Tr) regions are marked. Plasmid pAY includes 44 bp of the signal peptide (sp) of the Ehap-a gene. Both plasmids contain the ORF of Ehlgl1 gene and the 3′ regulatory sequence from the Ehactin gene.

(B) Northern blot analysis of amoebic RNA extracts. Lanes: 1, untransfected HM1:IMSS; 2, HM-1:IMSS transfected with plasmid pB33; 3, HM-1:IMSS transfected with pAY; 4, untransfected clone G3; 5, clone G3 transfected with pB33; 6, clone G3 transfected with pAY. Blots were probed as indicated.

(C) Western blot analysis of protein lysates separated on SDS-PAGE (12%) showing the blot that reacted with the polyclonal antibodies against the Lgl1 protein. Lanes 1–6 are as (B). Lane 7 contains lysate from the plasmidless RBV trophozoites (see Results).