Mechanisms of Functional Specificity Among Plasma-Membrane Syntaxins in Arabidopsis
Version of Record online: 28 JUN 2011
© 2011 John Wiley & Sons A/S
Volume 12, Issue 9, pages 1269–1280, September 2011
How to Cite
Reichardt, I., Slane, D., El Kasmi, F., Knöll, C., Fuchs, R., Mayer, U., Lipka, V. and Jürgens, G. (2011), Mechanisms of Functional Specificity Among Plasma-Membrane Syntaxins in Arabidopsis. Traffic, 12: 1269–1280. doi: 10.1111/j.1600-0854.2011.01222.x
- Issue online: 11 AUG 2011
- Version of Record online: 28 JUN 2011
- Received 20 April 2011, revised and accepted for publication 1 June 2011, published online 28 June 2011
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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