Fig. 6: mESCs display position-dependent differences in protein synthesis and proliferation rates.

a Representative images showing the protein synthesis of the micropatterned mESCs at 0, 12, 24 and 48 h after LIF withdrawal. Images from different time points are set as the same maximum and minimum grey value Scale bar, 120 µm. b Quantification of relative fluorescent intensity of the composite image of 2–9 micropatterns of the protein synthesis in mESCs at 0, 12, 24 and 48 h after LIF withdrawal (individual intensity profiles of micropatterns in Source Data). c Representative images showing the protein synthesis of the micropatterned mESCs at 48 h after LIF withdrawal (left). Scale bar, 120 µm. Quantification of relative fluorescent intensity of the protein synthesis in mESCs across 10 micropatterns at 48 h after LIF withdrawal (right). d Pie chart showing the percentage of mRNAs, proteins, and phosphoproteins of targets that are more abundant at the centre (purple) and targets that are more abundant at the edge (orange) of the micropattern. e, f Heatmap of DAPI intensity across the micropattern and normalised DAPI intensity across the micropattern at 0, 12, 24 and 48 h after LIF withdrawal. g Schematic shows the ‘donut’ shape of the micropatterned mESCs. h Representative images showing EdU staining of the micropatterned mESCs at 0, 12, 24 and 48 h after LIF withdrawal. Images from different time points are set as the same maximum and minimum grey value. Insets: an image of EdU staining at 48 h after LIF withdrawal with adjusted contrast. Scale bar, 120 µm. i Quantification of relative fluorescent intensity of EdU staining in mESCs of composite image of 3–10 micropatterns (individual intensity profiles of micropatterns in Source Data) at 0, 12, 24 and 48 h after LIF withdrawal. j, Heatmap of Ki-67 expression level at z-projection across 1 micropattern at 0, 12, 24 and 48 h after LIF withdrawal. Source data are provided as Source Data sheet tabs Fig. 6a–j.