Fig. 4: Microcontact-printed GFP and mCherry ligands spatially activate distinct reporter genes via synNotch in dual-reporter receiver fibroblasts. | Nature Communications

Fig. 4: Microcontact-printed GFP and mCherry ligands spatially activate distinct reporter genes via synNotch in dual-reporter receiver fibroblasts.

From: Engineering programmable material-to-cell pathways via synthetic notch receptors to spatially control differentiation in multicellular constructs

Fig. 4

A Schematic of dual-ligand microcontact printing and seeding of dual-receiver fibroblasts with anti-GFP/tTa synNotch that activates miRFP and orthogonal anti-mCherry/Gal4-VP64 synNotch that activates BFP. B Fluorescence microscopy images of microcontact-printed 500-μm-wide perpendicular lines of GFP and mCherry and corresponding miRFP and BFP expression by dual-receiver cells after one day of culture. Scale bars, 500 μm. Right: Profile plot of normalized miRFP intensity across the y-axis, green bars indicate regions patterned with GFP. The line represents mean ± s.d., n = 7 biological replicates. Below: Profile plot of normalized BFP intensity across the x-axis, red bars indicate regions patterned with mCherry. The line represents mean ± s.d., n = 7 biological replicates. The region outlined by the yellow rectangle is shown at a higher magnification below the image. Regions patterned with (i, red square) mCherry only, (ii, green square) GFP only, (iii, blue square) mCherry and GFP, and (iv, black square) neither mCherry nor GFP. C Higher magnification fluorescence microscopy images of regions shown in (B). Scale bars, 100 μm. D Percent of BFP and/or miRFP-expressing receiver cells after one day of culture on the indicated GFP/mCherry patterns, quantified by image analysis. n = 4 biological replicates, data represent mean ± s.d. See Fig. S5F, G for more data on this. Source data are provided as a Source Data file.

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