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. 2019 Jun 3;146(12):dev172940. doi: 10.1242/dev.172940

Fig. 4.

Fig. 4.

Analysis of ERK activity dynamics and Nanog expression during ESC differentiation after LIF removal. (A) Protocol for analysis of ERK activity dynamics during ESC cell differentiation following LIF removal. (B) Ratiometric FRET images of ESCs at indicated time points (min) following release from LIF followed by immunostaining for Nanog. Scale bar: 20 µm. (C) FRET/YFP levels of each cell (left) after LIF removal, pooled from five fields of view (n=303). Data from a typical experiment from three biological replicates (928 cells total). Mean FRET/YFP ratio of all cells shown on the right. See Fig. S4 for data from other replicates. (D) Mean (P=0.26), minimum (P=0.22) and maximum (P=0.32) ERK activity of each cell plotted against Nanog intensity from a typical replicate (190 cells).