Nova mlada zagrebačka scena devedesetih

In MVBs, the probe labelled delimiting (A, see arrow) and internal membranes (A, see arrowheads). Large tubular vesicles did not show particles (B). Negatively charged nanogold was also observed in Golgi bodies(C, see arrows). Bar=50 nm.<p><b>Copyright information:</b></p><p>Taken from "Clathrin-dependent and independent endocytic pathways in tobacco protoplasts revealed by labelling with charged nanogold"</p><p></p><p>Journal of Experimental Botany 2008;59(11):3051-3068.</p><p>Published online 4 Jul 2008</p><p>PMCID:PMC2504345.</p><p></p

(A) Controls of probe specificity: protoplasts incubated without nanogold were processed for silver enhancement and no dark spots were visible in the cells

(B, C) FDA test of protoplast viability after 120 min of incubation during the time-course experiments (B; M= 55X) and after 500 μM sodium azide treatment (C; M=68X). In all samples, protoplasts showed FDA-dependent fluorescence, indicating intact cell membranes and viable protoplasts. Bar=50 nm.<p><b>Copyright information:</b></p><p>Taken from "Clathrin-dependent and independent endocytic pathways in tobacco protoplasts revealed by labelling with charged nanogold"</p><p></p><p>Journal of Experimental Botany 2008;59(11):3051-3068.</p><p>Published online 4 Jul 2008</p><p>PMCID:PMC2504345.</p><p></p

Compartments labelled by positively charged nanogold and quantitation of the labelled profile during the Ikarugamycin experiment: protoplasts were incubated with IKA to inhibit clathrin-dependent endocytosis

(A) Nanogold accumulated on the plasma membrane and in some areas gold labelling formed clusters immediately below the plasma membrane (see arrows). (B, C) Some nanogold particles were observed in interconnected vesicle compartments (B, see arrow) and MVBs (C, see arrow). (D) Quantitation of particle distribution confirmed TEM observations. Since IKA was suspended in DMSO, the effects of IKA on nanogold internalization were compared with DMSO. IKA inhibited probe internalization and many particles were still found in inner vesicles after IKA treatment, suggesting that most of the traffic involving small inner vesicles was due to clathrin-independent processes bypassing tubular/round vesicles. Standard error (SE) is shown for quantitation data. Bar=100 nm.<p><b>Copyright information:</b></p><p>Taken from "Clathrin-dependent and independent endocytic pathways in tobacco protoplasts revealed by labelling with charged nanogold"</p><p></p><p>Journal of Experimental Botany 2008;59(11):3051-3068.</p><p>Published online 4 Jul 2008</p><p>PMCID:PMC2504345.</p><p></p

Quantitation of the labelled profile during energy-dependent experiments and temperature-dependent experiments

(A, B) For energy controls, protoplasts were pretreated and incubated with different concentrations of sodium azide. (A) At low sodium azide concentrations uptake of nanogold was not inhibited but differences in the distribution of the probe were observed with respect to the control. (B) At high concentrations of sodium azide entry of probe was inhibited. (C) For low temperature experiments, protoplasts were incubated at 4 °C for 30 min before addition of positively charged nanogold and samples were taken after incubation with the probe for 45 min at 4 °C. An increase in the number of particles was observed on the PM with respect to the control and a corresponding decrease of probe was observed in peripheral and large vesicles. (D) Recovery of intracellular trafficking after incubation at low temperature was performed by bringing to 25 °C and taking samples after 15 min and 45 min. Quantitation data suggested that, during recovery, internalization was not reactivated while the recycling pathways was restored. The increase of the probe in inner vesicles and in vacuoles was due to the recovery of vesicle trafficking. (E, F) Effect of sodium azide and low temperature on Golgi and ER labelling. Sodium azide and cold experiments showed that a very low level of labelling was observed in Golgi bodies and ER (E) and a low percentage of Golgi bodies were labelled (F) compared to the control. Standard error (SE) is shown for each experiments.<p><b>Copyright information:</b></p><p>Taken from "Clathrin-dependent and independent endocytic pathways in tobacco protoplasts revealed by labelling with charged nanogold"</p><p></p><p>Journal of Experimental Botany 2008;59(11):3051-3068.</p><p>Published online 4 Jul 2008</p><p>PMCID:PMC2504345.</p><p></p

Compartments labelled by positively charged nanogold during time-course experiments: protoplasts were incubated with 30 nmol of positively charged nanogold and fixed after 5, 15, 30, 45, and 120 min

(A) After 5 min of incubation most of the particles were distributed on the PM and peripheral vesicles (arrows). (B) Some large vesicles near the PM appeared labelled by the probe. (C, D) After 5–120 min of incubation, the probe was seen in small vesicles immediately below the PM (C, see blue arrowheads for peripheral vesicles and black arrowheads for PM) and into inner vesicles in the cortical region of cell (C, D, see black arrows) or deeper in the cytoplasm (C, D, see blue arrows). Golgi bodies appeared labelled by the probe (D, see arrowheads). (E) After 15–120 min of incubation, positively charged nanogold was seen inside large tubular membraneous compartments (large vesicles). (F) Some roundish-tubular vesicles showed internal membranes similar to MVBs labelled by the probe. (G, H, I) The ER (G, arrow) and Golgi bodies (H, I) also contained gold particles. (A, B, E, F, G, H, I) Bar=50 nm. (C, D) Bar=100 nm.<p><b>Copyright information:</b></p><p>Taken from "Clathrin-dependent and independent endocytic pathways in tobacco protoplasts revealed by labelling with charged nanogold"</p><p></p><p>Journal of Experimental Botany 2008;59(11):3051-3068.</p><p>Published online 4 Jul 2008</p><p>PMCID:PMC2504345.</p><p></p

Immunoblot and immunogold labelling by SYP21 polyclonal antibody, a marker of late endosomes

(A) The specificity of antibody was tested by western blot on tobacco protoplast crude extracts: lane CE showed SDS-PAGE of tobacco protoplast crude extract in which blot analysis using Syp 21 antibody (lane SYP21) specifically identified a single band at 30 kDa. Controls without primary antibody (lane C) did not show aspecific reactions. (B–D) In immunogold experiments, to distinguish the nanogold from gold-conjugated antibodies, the silver enhancement was performed for 1 min instead than 2 min. Nanogold particles were smaller than in the other experiments and smaller than the 20 nm gold particles bound to the secondary antibody. The antibody localized in interconnected vesicles (B; arrows) and MVBs (D, arrows) that were also labelled by nanogold (B, D, arrowheads). Multivesiculated vesicles fused with the central vacuole (asterisk). (C) Negative controls of immunogold analyses without primary antibody. No aspecific labelling was present. Bar=100 nm.<p><b>Copyright information:</b></p><p>Taken from "Clathrin-dependent and independent endocytic pathways in tobacco protoplasts revealed by labelling with charged nanogold"</p><p></p><p>Journal of Experimental Botany 2008;59(11):3051-3068.</p><p>Published online 4 Jul 2008</p><p>PMCID:PMC2504345.</p><p></p

Plant and arthropod colonisation of a glacier foreland in a peripheral mountain range

<div><p>Primary successions along glacier forelands are perfect examples of the changing climate upon high mountain ecosystems. Peripheral mountain ranges deserve particular attention, given they are characterised by high numbers of species and endemism and are considered to be particularly susceptible to climate change. We analysed thermal regime, soil parameters and plant/arthropod primary succession along a glacier foreland located in such a context, comparing it with those previously studied in the inner Alps. The overall patterns of the investigated primary succession agree with those of the inner Alps at the same elevation, but stands out for a delayed plant and arthropod colonisation which promotes the long-lasting persistence of pioneer cold-adapted species. In light of the results obtained, and considering the glaciological features of peripheral mountain ranges (glaciers persistence at low elevation), this paper asserts the hypothesis that glacial landforms of these areas may act as warm-stage refugia for pioneer cold-adapted species.</p></div

2D gel resolved seed polypeptides identified by MALDI-TOF/TOF MS.

<p>2D gel resolved seed polypeptides identified by MALDI-TOF/TOF MS.</p

2D gel electrophoresis of F18 seeds.

<p>Polypeptides changed in transgenic line with respect to the WT were evidenced by circle. The number of spots correspond to polypeptides identified by MALDI TOF/TOF MS analysis. Storage proteins are highlighted in black, Chaperone proteins in green, LEA proteins in violet, enzymes in blue, and other proteins in red. Spot numbers of the enhanced polypeptides compared to the WT are in bold.</p

1-D gel electrophoresis of dry and hydrated seeds crude extracts.

<p>Qualitative differences in the polypeptidic profile was observed: while the intensity of some polypeptides seemed to be equivalent in the WT and in transgenic seeds, others increased in transgenic lines (polypeptides 11–13, 14–15 and 16–18 in F18 dry seed and 19–21, 22–23, 24–26 in VT2eB dry seed). On the other hand, low molecular weight polypeptides were present only in WT seeds (bands 8–10 in dry seed). In hydrated seeds the difference in the polypeptidic profile was less pronounced, and only polypeptides with a molecular mass of 43 kDa, and comprised between 6.5 and 14 kDa were observed in the WT (asterisks). In moist WT seeds, the polypeptides corresponding to band 2 and those corresponding to bands 6–7 had a higher intensity than the dry WT seeds. The band numbers correspond to polypeptides identified by MALDI TOF/TOF MS analysis showed in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0187929#pone.0187929.t001" target="_blank">Table 1</a>. The band numbers of the enhanced polypeptides are in bold.</p