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.