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Commit 95839322 authored by Viktoria Petrova's avatar Viktoria Petrova
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fix assays' README

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![fpls-14-1182105-g002.jpg](dataset/fpls-14-1182105-g002.jpg)
Figure 2
**Figure 2**
Storage oil mobilization in castor bean endosperm. (A) Relative fresh weight of the endosperm in dry seeds (-1d), imbibed seeds (0d) and 1- to 7-day old dark-grown castor bean seedlings (1-7d) as percentage of fresh weight of the whole plant. Data represent arithmetic means ± SD of 3 biological replicates. (B) Relative ricinoleic acid content of the endosperm from seeds and seedlings. Samples were normalized to the average fresh weight of on endosperm and expressed as percentages of their initial quantities determined in dry seeds (-1d). Error bars show standard deviations of the means of at least three biological replicates.
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![fpls-14-1182105-g003.jpg](dataset/fpls-14-1182105-g003.jpg)
**Figure 3**
Isolation of organelles from etiolated castor bean seedlings. Four fractions of the sucrose density step gradient after centrifugation were taken from the gradient for various analyses at the interface 30% - 44% (w/w) sucrose solution (F1), 44% - 48% (w/w) sucrose solution (F2), 48% - 49% (w/w) sucrose solution (F3), and 50% - 54% (w/w) sucrose solution (F4).
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![fpls-14-1182105-t001.jpg](dataset\fpls-14-1182105-t001.jpg)
**Table 1**
Distribution of marker enzyme activities between the fractions.
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![fpls-14-1182105-g004.jpg](dataset\fpls-14-1182105-g004.jpg)
**Figure 4**
Purification of intracellular membranes from castor bean endosperm. Distribution of organellar membrane proteins within the organellar fractions (F1-F4) obtained after sucrose density gradient centrifugation. (A) SDS-PAGE analysis. Each lane was loaded with enriched membrane proteins from each pellet fraction after organelle disruption and ultracentrifugation (F1/F2: 25 µg, F3/F4: 15 µg). Equal loading of the protein gel within the fractions was monitored by staining with colloidal Coomassie G-250 dye. (B) Immunoblot analysis using antibodies raised against the following organelle membrane proteins: chloroplast TIC40, mitochondrial AOX, ER BIP2, and peroxisomal PEX14 proteins. Expected molecular weight for the corresponding castor bean proteins: 50.1 kDa for RcTIC40 (A3QSJ7), 39-41 kDa for RcAOX1/2 (B9RXE2/B9SO38), 73.3 kDa for RcBIP2 (B9RYP6), and 58.3 kDa for RcPEX14 (B9RB25). The membrane was serially probed with antibodies to the indicated marker proteins. The positions of molecular mass markers (in kDa) are indicated at the left.
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![fpls-14-1182105-g005.jpg](dataset\fpls-14-1182105-g005.jpg)
Source: https://www.frontiersin.org/articles/10.3389/fpls.2023.1182105/full#supplementary-material
**Figure 5**
Principal Component Analysis of the Ricinus proteome of the fractions obtained from the sucrose density centrifugation, representing the total proteins (T1 to T4) and the enriched membrane proteins (M1 to M4). In this PCA plot each point represents the identified proteome of one biological experiment.
![fpls-14-1182105-g006.jpg](dataset\fpls-14-1182105-g006.jpg)
**Figure 6**
Distribution profile of compartment-specific marker proteins in the density-gradient fractions containing the total proteins (T1 to T4) and membrane proteins (M1 to M4). (A) peroxisomes; (B) plastid; (C) mitochondria; (D) ER; (E) nucleus; (F) Golgi; (G) vacuole; (H) cytosol. Relative LFQ represents the sum of Label Free Quantification (LFQ) values for all proteins belonging to an organelle relative to its maximal value.
## Supplementary material
Source for supplementary material: https://www.frontiersin.org/articles/10.3389/fpls.2023.1182105/full#supplementary-material
## "Table 3.xlsx"
......@@ -11,7 +24,6 @@ Deconvolution-based assignment of the MS-identified proteins from etiolated cast
SUPPLEMENTARY TABLE 4
List of proteins found in peroxisomes, mitochondria, and plastids isolated from castor bean etiolated endosperm in this study. For functional annotation of proteins and their assignment to biological processes or metabolic pathways, gene ontology analysis was performed by Uniprot.
## "Image 1.jpg"
SUPPLEMENTARY FIGURE 1
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![fpls-14-1182105-g001.jpg](resources/fpls-14-1182105-g001.jpg)
Figure 1
**Figure 1**
Endosperm morphology in seeds and germinating seedling of R. communis. Photographs were taken from the whole plant (upper panel) and two cotyledons embedded in the endosperm (lower panel) from mature seeds (dry), 24h-imbibed seeds (imb) and from 1- to 7-day old dark-grown castor bean seedlings (1-7d). Scale bar = 1 cm.
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