Journal: bioRxiv

Article Title: A versatile method to expand and compare transporter substrate spectra

doi: 10.1101/2025.07.11.664331

Figure Lengend Snippet: Presence of indole-glucosinolates glucobrassicin (I3M), 4-methoxyglucobrassicin (4MOI3M), and neoglucobrassicin (NMOI3M) in oocytes exposed to 1:100 seedling media. NPF2.10 in brown, NPF2.11 in red, and NPF2.9 in pink. Oocytes were assayed at pH 5 for 1 hour; replicates consisted of five oocytes each, and media samples consisted of 5µl each. Control oocytes in blue and seedling media in green. Combined extracted ion chromatogram for I3M (447.0537 +/-0.005 m/z) and 4NMOI3M and NMOI3M (477.0643 +/-0.005 m/z).

Article Snippet: Neoglucobrassicin (NMOI3M), glucobrassicin (I3M), 4-methoxy-3-indolylmethyl glucosinolate (4MOI3M) sinalbin (pOHB), glucoraphanin (4msb) afzelin (K3R), isoquercetin (Q3G), quercitrin (Q3R), and astragalin (K3G) where purchased from Extrasynthese.

Techniques: Control

Journal: bioRxiv

Article Title: A versatile method to expand and compare transporter substrate spectra

doi: 10.1101/2025.07.11.664331

Figure Lengend Snippet: The metabolic differences of the extracts and transporter activity drive the chemical profile of the oocytes . Unsupervised bicluster analysis of oocytes assayed with empty buffer (pH5), and control (mock) and GTR-expressing oocyte samples assayed with either seed (Seed) or seedling (Sling) derived media. The relative intensities of metabolic features are represented on a color scale. Columns represent samples, and rows represent metabolic features. Oocytes were assayed at pH 5 for 1 hour with either 1:100 dilutions of seed or seedling extract; replicates consisted of five oocytes each. pH5 oocytes were assayed with an empty pH 5 buffer for 1 hour. Metabolic features identified with chemical standards as indole glucosinolates in black font: glucobrassicin (I3M), neoglucobrassicin (NMOI3M), and 4-methoxyglucobrassicin (4MOI3M). Annotated glucosinolates in blue font: 3-sinapoyloxypropylglucosinolate (3sin), 4-sinapoyloxybutylglucosinolate (4sin), glucoraphasatin (dH4mtb), 6′-O-benzoyloxy-glucoerucin (6’bz4mtb). Annotated flavonoid Quercitrin (Q3R) in purple. Ward’s biclustering of the 100 most significant features (ANOVA). Sig. P-value <0.05. Features displaying a Relative Standard deviation of >30% in the QC were filtered. Features are logarithmically transformed (Log10) and Pareto scaled. Identification of indole glucosinolate-derived features was performed with co-analyzed chemical standards.

Article Snippet: Neoglucobrassicin (NMOI3M), glucobrassicin (I3M), 4-methoxy-3-indolylmethyl glucosinolate (4MOI3M) sinalbin (pOHB), glucoraphanin (4msb) afzelin (K3R), isoquercetin (Q3G), quercitrin (Q3R), and astragalin (K3G) where purchased from Extrasynthese.

Techniques: Activity Assay, Control, Expressing, Derivative Assay, Standard Deviation, Transformation Assay

Journal: bioRxiv

Article Title: A versatile method to expand and compare transporter substrate spectra

doi: 10.1101/2025.07.11.664331

Figure Lengend Snippet: Glucosinolate import generates most of the clusters separating selected NPF substrate spectra, while phenylpropanoid import contributes to the separation of some additional samples . Unsupervised bicluster analysis of control (mock), and selected NPF-expressing oocyte samples assayed with 1:20 dilution of seedling-derived media. The relative intensities of metabolic features are represented on a color scale. Columns represent samples and rows metabolic features. Metabolic features identified with chemical standards are in black: glucobrassicin (I3M), neoglucobrassicin (NMOI3M), 4-methoxyglucobrassicin (4MOI3M), and Kaempferol-3-O-rhamnoside (K3R). Annotated glucosinolates in blue font: 3-sinapoyloxypropylglucosinolate (3sin), 4-sinapoyloxybutylglucosinolate (4sin), 4-Hydroxyglucobrassicin (4OHI3M), and 6′-O-benzoyloxy-4-sinapoyloxybutylglucosinolate (6’bz4sin). Features deriving from annotated phenylpropanoids in purple: Two isomers of disinapoyl hexose (DSH’’ and DSH’’’). Ward’s biclustering of the 100 most significant features (ANOVA). Sig. P-value <0.05. Oocytes were assayed at pH 5 for 1 hour with 1:20 dilutions of seedling extract; replicates consisted of five oocytes each. Features displaying a Relative Standard deviation of >30% in the QC were filtered. Features are logarithmically transformed (Log10) and Pareto scaled.

Article Snippet: Neoglucobrassicin (NMOI3M), glucobrassicin (I3M), 4-methoxy-3-indolylmethyl glucosinolate (4MOI3M) sinalbin (pOHB), glucoraphanin (4msb) afzelin (K3R), isoquercetin (Q3G), quercitrin (Q3R), and astragalin (K3G) where purchased from Extrasynthese.

Techniques: Control, Expressing, Derivative Assay, Standard Deviation, Transformation Assay