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. 2016 Oct 5;4:e2507. doi: 10.7717/peerj.2507

Figure 1. Ubi-XA21nls-GFP transgenic plants have reduced transcript and protein levels.

Figure 1

(A) Domain organization of XA21 receptor (SP, signal peptide; LRR, leucine rich repeat; TM, transmembrane domain; JM, juxtamembrane domain; KD, kinase domain). The predicted nuclear localization sequence (NLS: KRTKK, amino acids 678-682) highlighted in blue, is mutated to AATAA in Ubi-XA21nls-GFP. (B) Relative expression levels of XA21 in Ubi-XA21-GFP and T2 progeny derived from three independent Ubi-XA21nls-GFP T1 lines (XA21nls #1–8, #2–3 and #7–3) determined by qRT-PCR and normalized to ubiquitin reference gene. The XA21 expression level in Ubi-XA21-GFP was set to 1. Bars represent mean ± SD of three technical replicates. Different letters indicate significant differences between the groups (Tukey’s honestly significant difference test, α < 0.05). The experiment was repeated three times with similar results. (C) Western blot analysis of 70 µg total protein extracts from 5-wk Ubi-XA21-GFP, Kitaake and two individual T2 plants derived from three Ubi-XA21nls-GFP T1 lines. The full-length XA21-GFP was detected with an anti-GFP antibody. Equal loading of total proteins was confirmed by Coomassie blue staining (CBB).