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a , Schematic design of lipid transfer assay. Donor liposomes include NBD conjugated to phosphatidic acid (NBD-PA), whose fluorescence signal is quenched by FRET by rhodamine conjugated to phosphatidylethanolamine <t>(Rhod-PE).</t> Lipid transfer from donor liposomes to acceptor liposomes results in dequenching and increased fluorescence signal. b , SDS-PAGE gel of purified mSCP2. c – e , Fluorescence signal readout using donor liposomes with different POPA percentages ( c ), negative control where mSCP2 or acceptor liposomes were omitted ( d ), and with different mSCP2 concentrations ( e ). Raw fluorescence signal is shown except for e , where fluorescence signal was normalized to the No Acceptor control. The concentration of mSCP2 was 187.5 nM unless otherwise noted. Shown are representative plots from two independent experiments.
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a , Schematic design of lipid transfer assay. Donor liposomes include NBD conjugated to phosphatidic acid (NBD-PA), whose fluorescence signal is quenched by FRET by rhodamine conjugated to phosphatidylethanolamine <t>(Rhod-PE).</t> Lipid transfer from donor liposomes to acceptor liposomes results in dequenching and increased fluorescence signal. b , SDS-PAGE gel of purified mSCP2. c – e , Fluorescence signal readout using donor liposomes with different POPA percentages ( c ), negative control where mSCP2 or acceptor liposomes were omitted ( d ), and with different mSCP2 concentrations ( e ). Raw fluorescence signal is shown except for e , where fluorescence signal was normalized to the No Acceptor control. The concentration of mSCP2 was 187.5 nM unless otherwise noted. Shown are representative plots from two independent experiments.
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a , Schematic design of lipid transfer assay. Donor liposomes include NBD conjugated to phosphatidic acid (NBD-PA), whose fluorescence signal is quenched by FRET by rhodamine conjugated to phosphatidylethanolamine <t>(Rhod-PE).</t> Lipid transfer from donor liposomes to acceptor liposomes results in dequenching and increased fluorescence signal. b , SDS-PAGE gel of purified mSCP2. c – e , Fluorescence signal readout using donor liposomes with different POPA percentages ( c ), negative control where mSCP2 or acceptor liposomes were omitted ( d ), and with different mSCP2 concentrations ( e ). Raw fluorescence signal is shown except for e , where fluorescence signal was normalized to the No Acceptor control. The concentration of mSCP2 was 187.5 nM unless otherwise noted. Shown are representative plots from two independent experiments.
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a , Schematic design of lipid transfer assay. Donor liposomes include NBD conjugated to phosphatidic acid (NBD-PA), whose fluorescence signal is quenched by FRET by rhodamine conjugated to phosphatidylethanolamine <t>(Rhod-PE).</t> Lipid transfer from donor liposomes to acceptor liposomes results in dequenching and increased fluorescence signal. b , SDS-PAGE gel of purified mSCP2. c – e , Fluorescence signal readout using donor liposomes with different POPA percentages ( c ), negative control where mSCP2 or acceptor liposomes were omitted ( d ), and with different mSCP2 concentrations ( e ). Raw fluorescence signal is shown except for e , where fluorescence signal was normalized to the No Acceptor control. The concentration of mSCP2 was 187.5 nM unless otherwise noted. Shown are representative plots from two independent experiments.
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a , Schematic design of lipid transfer assay. Donor liposomes include NBD conjugated to phosphatidic acid (NBD-PA), whose fluorescence signal is quenched by FRET by rhodamine conjugated to phosphatidylethanolamine <t>(Rhod-PE).</t> Lipid transfer from donor liposomes to acceptor liposomes results in dequenching and increased fluorescence signal. b , SDS-PAGE gel of purified mSCP2. c – e , Fluorescence signal readout using donor liposomes with different POPA percentages ( c ), negative control where mSCP2 or acceptor liposomes were omitted ( d ), and with different mSCP2 concentrations ( e ). Raw fluorescence signal is shown except for e , where fluorescence signal was normalized to the No Acceptor control. The concentration of mSCP2 was 187.5 nM unless otherwise noted. Shown are representative plots from two independent experiments.
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a , Schematic design of lipid transfer assay. Donor liposomes include NBD conjugated to phosphatidic acid (NBD-PA), whose fluorescence signal is quenched by FRET by rhodamine conjugated to phosphatidylethanolamine (Rhod-PE). Lipid transfer from donor liposomes to acceptor liposomes results in dequenching and increased fluorescence signal. b , SDS-PAGE gel of purified mSCP2. c – e , Fluorescence signal readout using donor liposomes with different POPA percentages ( c ), negative control where mSCP2 or acceptor liposomes were omitted ( d ), and with different mSCP2 concentrations ( e ). Raw fluorescence signal is shown except for e , where fluorescence signal was normalized to the No Acceptor control. The concentration of mSCP2 was 187.5 nM unless otherwise noted. Shown are representative plots from two independent experiments.

Journal: Nature chemical biology

Article Title: Membrane editing with proximity labeling reveals regulators of lipid homeostasis

doi: 10.1038/s41589-025-02104-x

Figure Lengend Snippet: a , Schematic design of lipid transfer assay. Donor liposomes include NBD conjugated to phosphatidic acid (NBD-PA), whose fluorescence signal is quenched by FRET by rhodamine conjugated to phosphatidylethanolamine (Rhod-PE). Lipid transfer from donor liposomes to acceptor liposomes results in dequenching and increased fluorescence signal. b , SDS-PAGE gel of purified mSCP2. c – e , Fluorescence signal readout using donor liposomes with different POPA percentages ( c ), negative control where mSCP2 or acceptor liposomes were omitted ( d ), and with different mSCP2 concentrations ( e ). Raw fluorescence signal is shown except for e , where fluorescence signal was normalized to the No Acceptor control. The concentration of mSCP2 was 187.5 nM unless otherwise noted. Shown are representative plots from two independent experiments.

Article Snippet: The following lipids were purchased: DOPC (Avanti Polar Lipids, 850375), PtdIns(4,5)P 2 (Avanti Polar Lipids, 840046), DGS-NTA(Ni) (Avanti Polar Lipids, 790404), POPA (Avanti Polar Lipids, 840857), NBD-PA (Avanti Polar Lipids, 810176), NBD-PE (Avanti Polar Lipids, 810144), Rhod-PE (Avanti Polar Lipids, 810150), DOPE (Cayman, 15091) and DOPC (Echelon, L-1182).

Techniques: Liposomes, Fluorescence, SDS Page, Purification, Negative Control, Control, Concentration Assay