Supplementary Figure 3: Bi-allelic up-regulation of Xist is reversible. | Nature Structural & Molecular Biology

Supplementary Figure 3: Bi-allelic up-regulation of Xist is reversible.

From: A symmetric toggle switch explains the onset of random X inactivation in different mammals

Supplementary Figure 3

(a) Simulation of doxycycline treatment one day before the onset of differentiation (linked to Fig. 4a–c). Boxplots show the frequency of mono-allelic (left) and bi-allelic Xist expression (right) in dox-treated (grey) and control cells (black) for 100 parameter sets that could reproduce mono-allelic Xist up-regulation. (b) Boxplots show the simulation results for artificial bi-allelic Xist induction as described in Fig. 4e in the main text, using the same parameters sets as in (a). On each box, the central mark indicates the median, and the bottom and top edges of the box indicate the 25th and 75th percentiles, respectively. The whiskers extend to the most extreme data points not considered outliers, and the outliers are plotted individually. (c, d) Bi-allelic Xist up-regulation is artificially induced by treating TX1072dT cells with doxycycline after 48h of differentiation (cp. Fig. 4e). Cells were treated with EdU to assay proliferation through measuring its incorporation into DNA during replication. EdU was labeled fluorescently through Click-it chemistry and Xist was visualized by RNA FISH. (c) The EdU-positive fraction was quantified at the indicated time points within cells expressing Xist mono- (black) and bi-allelically (grey). Mean and s.d. of n=3 independent experiments are shown, in each replicate at least 50 cells were counted per group, except for bi-allelic cells at 48h. * paired two-sample two-sided T-test. Scale bar indicates 5 μm.

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