Fig. 7

Clone interactions among parental and clonal cell lines in co-culture for 28 days (a-b). a Expected population percentage of total population calculated using cumulative population doublings (CPDs) per day vs. observed population percentage. b CPDs/day of single culture vs. CPDs/day of the same cell line in co-culture. c-e Cooperation of GFP C3 and mKO E10 in invasion assays: (c-d) Evaluation of cooperation on the invading capacity of single clones vs co-cultured clones: (c) Total number of invading cells and (d) Relative ratio of invading cells based on the cell number per clone cultured individually. (e) Invading capacity of cells conditioned with their own medium or medium from another clonal cell line. Relative ratio of invading cells based on the cell number per clone cultured with their own conditioned medium. (f-i) In vivo clonal cooperation. Cooperation of GFP C3 and mKO E10 in in vivo migration assays (Zebra fish model). Disseminated cells from the duct of Cuvier to the tail 72 h after injection: individual injection of GFP C3 and mKO E10 and co-injection of both clonal cell lines: (f) Total number of disseminated cells, (g) representative graph (scale bar = 70 μm), (h) relative ratio of invading cells based on the initial cell number per clone, (i) analysis of the coexistence of clones in the co-injected fish: percentage of co-injected fish with GFP C3 cells only, mKO E10 cells only, or both. Significant differences were determined using unpaired t-test with Welch’s correction. Asterisks indicate significant differences when P-values are < 0.05 (*), < 0.01 (**), and < 0.001 (***)