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. 2022 Mar 14;10(2):e00002-22. doi: 10.1128/spectrum.00002-22

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Copyright © 2022 Ranjit et al.

This is an open-access article distributed under the terms of the Creative Commons Attribution 4.0 International license.

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FIG 3 — Fluorescence-based sphingolipid kinase assay using NBD-labeled lipid substrates. (A) Expression of PG1348 from pET24b in E. coli analyzed by protein gel electrophoresis. Lanes: 1st, protein ladder; 2nd, empty vector pET24B; 3rd and 4th, pET24b-PG1348 uninduced and induced with 10 μM IPTG, respectively. (B) NBD-sphingosine assay on cell lysates from E. coli. (C) NBD-dihydrosphingosine assay on cell lysates from E. coli. (D) NBD-dihydrosphingosine assay on cell lysates from the parent strain W83 and the W83 ΔPG1348 mutant. Number of repeats (n), 3. Error bars shown in the graph were calculated using the following formula: SE = SD/√n, where SE is the standard error of the sample, SD is the standard deviation of the sample, and n is the number of samples.