Supplemental Data

Files in this Data Supplement:

• Supplemental Figure 1 - Detergent Resistant Components in Bienertia sinuspersici and Suaeda aralocaspica Chlorenchyma Cells. Scanning electron micrographs of Bienertia (A-C) and S. aralocaspica (D-G) chlorenchyma cells treated in detergent (Triton X-100) depicting detergent resistant components (arrowheads) associating with chloroplasts within the cells. Bars in A = 15 μm, D, E, and F = 5 μm, and B, C, and G = 1 μm.
• Supplemental Figure 2 - Specificity of the Anti-Actin and Anti-Tubulin Antibodies. Immunoblot analysis showing the specificity and reactivity of anti-actin and anti-β-tubulin antibodies with Bienertia and S. aralocaspica total protein. Ten μg of total protein from Bienertia (lanes 2 and 4) and S. aralocaspica (lanes 3 and 5) mature leaves were probed with antibodies raised against human skeletal muscle actin and rat brain β -tubulin. Numbers on the left indicate molecular mass in kDa (lane 1).
• Supplemental Figure 3 - The Actin and Microtubule Cytoskeleton in Chlorenchyma Cells of Suaeda heterophylla (C₃) and Suaeda eltonica (C₄ Kranz). Immunofluorescence staining of actin filaments (A, B, E, and F) and microtubules (C, D, G, and H) demonstrating actin- and microtubule-chloroplast association in S. heterophylla (A-D) and S. eltonica (E-H) chlorenchyma cells, respectively. Actin filaments and microtubules (green) were visualized with Oregon-green conjugated secondary antibody and chloroplasts (red) were observed using their autofluorescence. The images represent merged images of the dual channels which show their interaction. This is a representative result from at least three independent experiments with more than thirty cells observed. (A) and (C) Projections of low resolution images demonstrating the overall actin and microtubule cytoskeleton in mesophyll cells of S. heterophylla, respectively. (B) and (D) Optical sections of high magnification images showing the direct interaction between actin filaments and microtubules and chloroplasts in mesophyll cells of S. heterophylla. (E) and (G) Projections of low resolution images showing the actin and microtubule cytoskeleton in mesophyll cells of S. eltonica, respectively. (F) and (H) Projections of low resolution images showing the actin and microtubule cytoskeleton in bundle sheath cells of S. eltonica, respectively. The insets in (E) to (H) are high resolution optical sections showing the direct interaction between the cytoskeleton and chloroplasts in S. eltonica. Bars in A, C, E, F, and H = 10 μm; Bars in B, D, and insets = 5 μm; Bar in G = 15 μm.
• Supplemental Figure 4 - Effect of Cytochalasin D (CD) and Oryzalin (Ory) on Bienertia Chlorenchyma Cell Viability. Drug-treated chlorenchyma cells were rinsed in PME buffer containing 0.5 M mannitol and incubated in the same buffer containing the appropriate concentrations of rhodamine 123 and DIOC₆(3) 24 h after removal of the drugs. (A), (B), and (C) Single optical sections through the mid section of control (A), CD- (B) or Ory-treated (C) chlorenchyma cells stained with rhodamine 123. Only control and CD-treated chlorenchyma cells showed the concentration of fluorescently stained mitochondria (yellow-green) in the CCC. Note the lack of CCC organization and staining in Ory-treated cells. (D), (E), and (F) Projections of control (D), CD- (E) or Ory-treated (F) chlorenchyma cells stained with DiOC₆ (3). Staining of membranous structures (yellow-green) was observed only in control and CD-treated chlorenchyma cells. Note the absence of staining in Ory-treated cells. Bar = 20 μm.