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. 2011 Mar 12;39(13):5538–5554. doi: 10.1093/nar/gkr124

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© The Author(s) 2011. Published by Oxford University Press.

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 3. — Inhibition of L1 retrotransposition by rabbit A1 with catalytic site mutations. (A) 293T cells were co-transfected with 0.5 µg of the expression plasmids for rabbit A1 catalytic site mutants and 1.5 µg of pL1_RP-EGFP. GFP expression within the transfected 293T cells were analyzed on flow cytometry as described. Relative retrotransposition frequency in the absence of APOBEC proteins (vector) was set as 1.0. The histogram bars represent the mean of three independent cultures, and the standard deviation is shown. (B) Western blot analysis was performed by using extracts from 293T cells transfected by the expression plasmids for rabbit A1 catalytic site mutants and detected by using antibodies specific for the epitopes present in the test proteins. (C) The transposition frequency of L1 in 293T cells co-transfected with 1.5 µg of pL1_RP-EGFP along with decreasing amounts (0.5, 0.25 and 0.125 µg) of the expression plasmids for rabbit A1 catalytic site mutants as described in (A). The histogram bars represent the mean of three independent cultures, and the standard deviation is shown. (D) Western blot analysis of the protein expression levels from the experiment in (C).