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. 2015 Jul 2;43(14):7152–7161. doi: 10.1093/nar/gkv655

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© The Author(s) 2015. Published by Oxford University Press on behalf of Nucleic Acids Research.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted reuse, distribution, and reproduction in any medium, provided the original work is properly cited.

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Figure 4. — Comparison of single and dual regulation by chemical induction. (A) Representative wild-type Nicotiana benthamiana leaves were spot infiltrated to compare the extent of HR phenotype from (i, ii) pTA7001 avrBs2-HA (only regulated by the Dex-inducible promoter) or (iii, iv) pTA7001 avrBs2-HyP5SM-HA (also includes splicing regulation), with or without 30 μM Dex treatment. Leaf spots were also co-infiltrated with pTA7001 OsL5–6xHis and p1776 Bs2-HA. (B) Top: RT-PCR detection of avrBs2-HyP5SM-HA transcripts from transiently transformed N. benthamiana leaves at different time points post induction with Dex. Controls shown are no RNA or no RT enzyme in cDNA synthesis reaction and RT-PCR from wild-type tissue. Bottom: RT-PCR detection of OsL5 transcripts. (C) Top: Anti-HA western blot to detect protein expression from avrBs2-HA or avrBs2-HyP5SM-HA constructs induced with Dex or mock induced. Samples are from biological replicates. Leaf tissue were also co-infiltrated with pTA7001 OsL5–6xHis, except for the wild-type control. Middle: Anti-OsL5 western blot. Bottom: Ponceau S stain of the large subunit of RuBisCo as loading control.