Fig. 1. Structure and sequence of FLN1, HSA1 and TRXz. A, Abbreviated diagram of TRXz, HSA1 and FLN1 protein sequences. B, Amino acids sequences of the three proteins. Sequences of the chloroplast transit peptide (CTP) are shown in red bold type. The arrows indicate the putative cleavage site during chloroplast import based on the TargetP website. C, cDNA sequences encoding the three genes. Sequences of the three CTPs are shown in blue bold type.

Fig. 2. Subcellular localization of TRXz and other proteins.A-E, Diagrams of vector construction; F, The free green fluorescent protein (GFP) protein as a control. G, The ΔCTPTRXz protein localized to the cytoplasm. H, The only N-terminal 56 amino acids of the TRXz and GFP fusion protein localized to chloroplast. I-K, The CTPTrxz-Ghd10, CTPTrxz-MFS1 and CTPTrxz-SUI1 fusion proteins localized to the chloroplasts. All of the GFP fusion proteins carried a C-terminal GFP tag. In F-K, GFP fluorescence, chlorophyll autofluorescence, merged GFP and chlorophyll fluorescence, and bright-field images are shown. Bar = 5 μm.

Fig. 3. Subcellular localization of HSA1 and other proteins.A-D, Diagrams of vector construction. E, The free green fluorescent protein (GFP) protein as a control. F, The only N-terminal 54 amino acids of the HSA1 and GFP fusion protein localized to chloroplast. G-I, The CTPHSA1-Ghd10, CTPHSA1-MFS1 and CTPHSA1-SUI1 fusion proteins localized to the chloroplasts. All of the GFP fusion proteins carried a C-terminal GFP tag. In E-I, GFP fluorescence, chlorophyll auto fluorescence, merged GFP and chlorophyll fluorescence, and bright-field images are shown. Bar = 5 μm.

Fig. 4. Subcellular localization of FLN1 and other proteins. A-E, Diagrams of vector construction. F, The free green fluorescent protein (GFP) protein as a control. G, The Δ36CTPFLN1 protein localized to the cytoplasm. H, The only N-terminal 36 amino acids of the FLN1 and GFP fusion protein localized to chloroplast. I-K, The 36CTPFLN1-Ghd10, 36CTPFLN1-MFS1, and 36CTPFLN1-SUI1 fusion proteins localized to the chloroplasts. All of the GFP fusion proteins carried a C-terminal GFP tag. In F-K, GFP fluorescence, chlorophyll auto fluorescence, merged GFP and chlorophyll autofluorescence, and bright-field images are shown. Bar = 5 μm.

Fig. 5. Subcellular localization of other FLN1-related proteins. A-D, Diagrams of vector construction. E, The free green fluorescent protein (GFP) protein as a control. F, The Δ50CTPFLN1 protein localized to the cytoplasm. G, The Δ89CTPFLN1 protein localized to the cytoplasm. H and I, The 50CTPFLN1-SUI1 and 89CTPFLN1-SUI1 fusion proteins localized to the chloroplasts. All of the GFP fusion proteins carried a C-terminal GFP tag. In E-I, GFP fluorescence, chlorophyll autofluorescence, merged GFP and chlorophyll fluorescence, and bright-field images are shown. Bar = 5 μm.