Supporting Information

Additional Supporting Information may be found in the online version of this article:

Figure S1: Subcellular localization of PEN1 and SYP132 during cytokinesis. A–F) Like KNOLLE (A, D), MYC-PEN1 (B) and MYC-SYP132 (E) accumulate at the cell plate (arrows). MYC-PEN1 accumulates less at the plasma membrane, whereas MYC-SYP132 shows no difference (arrowheads). C and F) Overlay counterstained with DAPI (DNA, blue). G and H) Quantitative scans of MYC-PEN1 (G) and MYC-SYP132 (H) protein accumulation from the plasma membrane (PM) across the cell including the cell plate (CP). Scale bars, 5 µm.

Figure S2: Quantification of RFP-SYP1 protein accumulation at infected leaf cells. A–C, upper panels) UBQ:RFP-PEN1 and (D–F, upper panels) UBQ:RFP-SYP132 label the plasma membrane and the B. g. hordei–Arabidopsis interaction sites. Scale bars, 5 mm. A–F, lower panels) Quantitative scans of RFP-SYP1 protein accumulation from the plasma membrane across the cell to the penetration site. G) Quantification of signal intensity of the RFP-SYP1 proteins at the penetration site compared to the plasma membrane. n = 5; error bars indicate standard deviation.

Figure S3: Co-labeling of GFP-PEN1 and RFP-SYP132. A–C) Subcellular localization of GFP-PEN1 (A) and RFP-SYP132 (B) in Arabidopsis root-tip cells. Both SYP1 proteins label the plasma membrane (arrows), whereas only PEN1 labels some endosomes (arrowheads); (C) merged image. D–F) After BFA treatment PEN1 (D) but not SYP132 (E) accumulates in BFA compartments; (F) merged image. Scale bars, 5 µm.

Figure S4: Subcellular behavior of SYP1 syntaxins. A–C) Like KNOLLE (A), MYC-PEN1 (B, red) and MYC-SYP132 (C, red; arrowhead) accumulate in BFA compartments in mitotic cells. Only PEN1 accumulates in BFA compartments in interphase cells (B). D–K) Immuno-localization of RFP-PEN1 (D–G) and RFP-SYP132 (H–K) expressed from the HISTONE 4 (H4) promoter. D–F) RFP-PEN1 (red) localizes at the plasma membrane and colocalizes with the TGN/endosomal marker ARF1 (green). G) RFP-PEN1 accumulates in BFA compartments. H–J) RFP-SYP132 (red) localizes at the plasma membrane but does not colocalize with ARF1 (green). K) KNOLLE (green) but not RFP-SYP132 (red) accumulates in BFA compartments (arrowhead). Scale bars, 5 µm.

Figure S5: Protein levels of PEN1-KNOLLESND. Protein expression of transgenic PEN1-KNOLLESND from several independent transgenic lines was analyzed with anti-RFP antibody. Note that protein levels are nearly equal when expressed from the KNOLLE (KN) promoter or from the HISTONE 4 (H4) promoter. KN:RFP-PEN1-KNOLLESND transgenic lines #2 and #4 were shown to rescue knolle mutants like wild type (Table S1).

Table S1: Rescue analysis of SYP1 syntaxins. Progeny of transgenic plant lines were grown on agar plates to seedling stage and phenotypically analyzed. knolle and partial rescue phenotypes were counted.

Table S2: Oligonucleotide sequences.

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TRA_1222_sm_tS1_tS2.pdf203KSupporting info item

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