Fig. 1.

Expression and Solubility of GFP N-RICs and C-RICs. General schematic of the design of RIC with antigen fused at the C-terminus (C-RIC) or N-terminus (N-RIC) of the 6D8 IgG heavy chain. The RIC binds to the epitope tag of other RIC molecules, forming large immune complexes. Constructs N-H and C-H lack the 6D8 light chain, while construct C-HL lacks the epitope tag. (B,C) GFP was incorporated into N-RIC, C-RIC, or the variants described above and expressed in the leaves of N. benthamiana. Leaves were photographed under UV light at 5 DPI and a representative image is shown. (D) Clarified protein extracts from leaf spots agroinfiltrated with GFP C-RIC, N-RIC, or C-HL were separated by SDS-PAGE and imaged to show total protein or UV fluorescent protein bands. (E) The relative GFP expression of C-RICs, N-RICs, and C-HL was analyzed by band densitometry using endogenous plant protein bands as an internal loading control. Columns represent mean ± the standard error of 3 independently infiltrated leaf samples. The fluorescence intensity of C-HL was arbitrarily defined as 1. The p values were measured by student’s t-test.