JCS012138 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig S1. (A) Sections of pollen tubes not previously incubated with the probes were processed for silver enhancement. A few particles were occasionally observed on the cell wall of the granule (A,b), but no staining was seen in the cytoplasm (A,a,c). (B) To avoid crosstalk between the two probes control experiments were performed in which pollen tubes were loaded with BODIPY-Ceramide only (a,a′) and images were taken using the parameters used for the FM4-64. A very low level of background was observed (a′′) so excluding artefacts.

• Supplemental Figure S2 -

  Fig S2. In order to determine the real diameter of the probes, EM observations were carried out to measure the positively and negatively charged nanogold. The size of particles was variable, mostly ranging from 1.29-1.89 and 1.29-1.60 for positively (a,b) and negatively (a′,b′) charged nanoparticles.

• Movie 1 -

  Movie 1. Uptake of FM4-64 in actively growing pollen tubes of Nicotiana tabacum. As the PM becomes stained the fluorochrome internalization is fast. The entrance in the subapical regions is particularly evident.

• Movie 2 -

  Movie 2. Internalized membranes are accumulated in the tip region, particularly at the site of tube growth. The movie shows the dynamics of FM4-64-labelled membranes during changes in the growth direction of the pollen tube.

• Movie 3 -

  Movie 3. Uptake of FM4-64 pollen tubes of Nicotiana tabacum treated with 3 μM Ika. The PM becomes stained and the fluorescence is more pronounced in the subapical regions of the PM. However, the fluorochrome uptake is low compared to the control (Movie 1).