Supplementary Figure 5: Characterization of HES3 MIXL1^GFP/+ cells exposed to a localized BMP4 source in a smaller cell chamber

(a) Design of the two layers for the SU8 mold of the device with 500μm-diameter cell chambers; detail of a single unit. In the final device the white parts will be occupied by the PDMS and the colored parts will be empty. (b) Computational simulation of the diffusion of a reference molecule from the source side of the 500μm-diameter cell chamber after 48 hours of perfusion. (c) Calculated distribution of the reference molecule along the red line reported in Supplementary Fig. 5b at the indicated time points. (d) Fluorescence pictures for indicated markers in HES3 MIXL1^GFP/+ cells after 48 hours of exposure to a localized 40ng/ml source of BMP4 in the 500μm-diameter chambers microfluidic device. This experiment was repeated independently two times and showed similar results. (e) Mean (line) and standard error of the mean (light-colored area) of normalized fluorescence intensity for the indicated markers along the source-sink axis of multiple cell chambers of HES3 MIXL1^GFP/+ cells after 48 hours of exposure to a localized 40ng/ml BMP4 source in 500μm-diameter chambers (n=8 independent samples).