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Expi293F cells co-expressing wildtype or variants of myc-tagged AtSWEET13 and sucrose FRET biosensor, <t>FlipSuc-90u</t> delta1, were incubated in Hank’s buffered saline solution, pH 7.4 (HBSS). At time point 40s, 40 mM sucrose in HBSS was added, and a negative ratio change was observed. Final solution concentration is equivalent to 20 mM sucrose. For the negative control, no significant change in the ratio was observed since cells expressed only the sucrose biosensor. Data are mean ± SEM, N = 3 biological replicates.
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Expi293F cells co-expressing wildtype or variants of myc-tagged AtSWEET13 and sucrose FRET biosensor, <t>FlipSuc-90u</t> delta1, were incubated in Hank’s buffered saline solution, pH 7.4 (HBSS). At time point 40s, 40 mM sucrose in HBSS was added, and a negative ratio change was observed. Final solution concentration is equivalent to 20 mM sucrose. For the negative control, no significant change in the ratio was observed since cells expressed only the sucrose biosensor. Data are mean ± SEM, N = 3 biological replicates.
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Expi293F cells co-expressing wildtype or variants of myc-tagged AtSWEET13 and sucrose FRET biosensor, <t>FlipSuc-90u</t> delta1, were incubated in Hank’s buffered saline solution, pH 7.4 (HBSS). At time point 40s, 40 mM sucrose in HBSS was added, and a negative ratio change was observed. Final solution concentration is equivalent to 20 mM sucrose. For the negative control, no significant change in the ratio was observed since cells expressed only the sucrose biosensor. Data are mean ± SEM, N = 3 biological replicates.
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Expi293F cells co-expressing wildtype or variants of myc-tagged AtSWEET13 and sucrose FRET biosensor, <t>FlipSuc-90u</t> delta1, were incubated in Hank’s buffered saline solution, pH 7.4 (HBSS). At time point 40s, 40 mM sucrose in HBSS was added, and a negative ratio change was observed. Final solution concentration is equivalent to 20 mM sucrose. For the negative control, no significant change in the ratio was observed since cells expressed only the sucrose biosensor. Data are mean ± SEM, N = 3 biological replicates.
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Expi293F cells co-expressing wildtype or variants of myc-tagged AtSWEET13 and sucrose FRET biosensor, <t>FlipSuc-90u</t> delta1, were incubated in Hank’s buffered saline solution, pH 7.4 (HBSS). At time point 40s, 40 mM sucrose in HBSS was added, and a negative ratio change was observed. Final solution concentration is equivalent to 20 mM sucrose. For the negative control, no significant change in the ratio was observed since cells expressed only the sucrose biosensor. Data are mean ± SEM, N = 3 biological replicates.
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Expi293F cells co-expressing wildtype or variants of myc-tagged AtSWEET13 and sucrose FRET biosensor, <t>FlipSuc-90u</t> delta1, were incubated in Hank’s buffered saline solution, pH 7.4 (HBSS). At time point 40s, 40 mM sucrose in HBSS was added, and a negative ratio change was observed. Final solution concentration is equivalent to 20 mM sucrose. For the negative control, no significant change in the ratio was observed since cells expressed only the sucrose biosensor. Data are mean ± SEM, N = 3 biological replicates.
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Expi293F cells co-expressing wildtype or variants of myc-tagged AtSWEET13 and sucrose FRET biosensor, FlipSuc-90u delta1, were incubated in Hank’s buffered saline solution, pH 7.4 (HBSS). At time point 40s, 40 mM sucrose in HBSS was added, and a negative ratio change was observed. Final solution concentration is equivalent to 20 mM sucrose. For the negative control, no significant change in the ratio was observed since cells expressed only the sucrose biosensor. Data are mean ± SEM, N = 3 biological replicates.

Journal: bioRxiv

Article Title: Deep mutational scanning reveals sequence to function constraints for SWEET family transporters

doi: 10.1101/2024.06.28.601307

Figure Lengend Snippet: Expi293F cells co-expressing wildtype or variants of myc-tagged AtSWEET13 and sucrose FRET biosensor, FlipSuc-90u delta1, were incubated in Hank’s buffered saline solution, pH 7.4 (HBSS). At time point 40s, 40 mM sucrose in HBSS was added, and a negative ratio change was observed. Final solution concentration is equivalent to 20 mM sucrose. For the negative control, no significant change in the ratio was observed since cells expressed only the sucrose biosensor. Data are mean ± SEM, N = 3 biological replicates.

Article Snippet: The gene encoding the sucrose biosensor for fluorescence resonance energy transfer (FRET), FlipSuc-90u delta1, was subcloned from pRSET FlipSuc-90u delta1 into the BamHI-HindIII sites of pcDNA3.1(+). pRET FlipSuc-90u delta1 was a gift from Wolf Frommer (Addgene plasmid #14475; http://n2t.net/addgene:14475 ; RRID: Addgene_14475).

Techniques: Expressing, Incubation, Saline, Concentration Assay, Negative Control