Figure 1. Selection of mVenus+ transformants
by flow cytometry.
A. Heatmap representation of the % mVenus+ cells in
each transformant, as assessed by flow cytometry. mVenus fluorescence
intensity was acquired on the FL1 channel (filter at 525/10 nm) on 384
transformed antibiotic resistant microalgae transferred to liquid TAP
medium in four 96 well plates for 6 days. Negative controls (WT) are
represented in wells 95 and 96 of each plate. Crossed wells are
transformant that did not grow in liquid medium. B. Gating strategy and
representative dot plots of a negative (left panel) and a positive
(right panel) sample. C. Correlation between mean fluorescence intensity
and % of mVenus+ cells in 23 mVenus+ transformants
and a wild type strain re-assessed for mVenus production 6 weeks
following the first screening.
Figure 2. mVenus expression following treatment with
silencing inhibitors.
A. Heatmap representations of the % of mVenus expressing cells (top)
and of the mVenus delta mean fluorescence intensity (bottom) in clones
16, 18, 20, 21, 22 and WT. Liquid culture in 96 wells were treated with
DMSO (0.1%), Sirtinol (SIRT, 50 µM), vorinostat (SAHA, 2.5 µM),
OSS_128167 (OSS, 100 µM), nicotinamide (NAD, 1mM), Sodium butyrate
(ButNa, 5 mM), or a mix of all HDACi (Mix) for 24 hours. B.
Representative pseudocolor dot plots of mVenus levels in transformants
treated with HDACi or DMSO. *: p>0.05; **
p>0.01, using one-way ANOVA Friedman’s test with repeated
measures.