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. 2017 Dec 1;130(23):4002–4012. doi: 10.1242/jcs.208975

Fig. 3.

Fig. 3.

The cytoplasmic domain of TgrB1 is required for cooperative aggregation and differentiation. (A–C) Ovoids represent cells, protrusions represent TgrB1 and TgrC1 proteins, and colors represent allotypes: AX4 (tan) and tgrB1C1QS31 (red). Black protrusions represent extra alleles: tgrB1AX4-HA (green) and tgrB1AX4Δ828-902-HA (purple). Cooperative aggregation of mixes of 90% unlabeled AX4 cells with 5% GFP-labeled AX4 and 5% RFP-labeled tgrB1C1QS31tgrB1AX4-HA (B) or tgrB1C1QS31tgrB1AX4Δ828-902-HA (C). The spatial distribution variance (V) is shown inside each frame. A comparison between the variances is shown below the images with a P-value calculated from F-tests; *P<0.000001. Scale bar: 0.3 mm. (D) Cooperative differentiation of mixes of 0.2% cells carrying cotB-GFP with 99.8% unlabeled AX4 cells. Protrusions represent TgrB1 and TgrC1 and the colors represent alleles as above. Full green arrow inside the cells shows cotB-GFP expression; empty arrow indicates no expression. We evaluated GFP fluorescence levels by flow cytometry and plotted the results as histograms. The x-axis represents fluorescence intensify (arbitrary units) and the y-axis the number of events. The black bar indicates the GFP-positive populations and the numbers indicate the respective fractions (%) of GFP-positive cells.