Figure 4

Lent-On-Plus (CEETln2 and CEETln2Is2) LVs efficiently generates Doxycycline-responsive hESCs without selection or cloning.

(a) Representative plots showing eGFP expression profiles of hESCs (AND-1) control (Mock) and hESCs transduced with CESTnl2, CESTnl2Is2, CEETnl2 and CEETnl2Is2 LVs in the absence (top) or presence (bottom) of Dox (0.1 μg/ml). hESCs were transduced at MOI = 5 with the different LVs and analyzed 10 days later. The percentages (%) of the eGFP+ populations are shown in each plot. The MFIs of the whole living cells (WLC) are indicated at the bottom of each plot. The vcn/c of transduced cells are shown at the bottom of the figure. (b) Graphs showing fold induction (left) and leaking (right) of the CEETnl2 and CEETnl2Is2 LVs in hESCs at different time points during expansion (day 5, day 12 and day 33 post-transduction). Values represent mean +/− standard error of the mean of at least three separate experiments using AND-1 and H9 hESCs transduced at MOI = 5 with the different LVs (*p < 0.05). (c) Graph showing relative eGFP expression levels of CEETnl2 and CEETnl2Is2-transduced hESCs (AND-1) upon withdrawal (day 0) and addition (Day 12) of Dox along time in culture. Data represent fluorescence intensity relative to Day 0. Values represent mean +/− standard error of the mean of at least three separate experiments.