![Figure 8. Synergistic effects of combinatorial expression of nectins and cadherins on asymmetric mosaic cellular patterning. (A) The mosaic-forming assay using neuro-2a (n2a) cells expressing different combinations of cadherins and nectins. Boundaries between the combinations of n2- and n2-n2a cells (left), n2- and n3-n2a cells (left middle), [n2+Ecad]- and [n2+Ecad]-n2a cells (right middle), and [n2+Ecad]- and [n3+Ecad]-n2a cells (right) are indicated. n2-n2a, neuro-2a nectin-2 transfectants; n3-n2a, nectin-3 transfectants; [n2+Ecad]-n2a, nectin-2 and E-cadherin transfectants; [n3+Ecad]-n2a, nectin-3 and E-cadherin transfectants. The graph shows the number of singly isolated cells in the mixtures of the two cell populations in each field. The results shown are the mean ± SD; *, P < 0.001; n.s., not significant. n = 9. (B) The mosaic-forming assay of HEK293 (293) cells expressing different cadherins and nectins. Boundaries between the combinations of n2- and n2-293 cells (top), n2- and n3-293 cells (middle), and n2- and [n3+Ecad]-293 cells (bottom) are indicated. White dotted lines indicate the center of the field. n2-293, 293 nectin-2 transfectants; n3-293, nectin-3 transfectants; [n2+Ecad]-293, nectin-2 and E-cadherin transfectants; [n3+Ecad]-293, nectin-3 and E-cadherin transfectants. (C) The graph indicates the number of singly isolated cells in the counter colony in each field. The results shown are the mean ± SD; *, P < 0.05. n = 9. (D) Distribution patterns of β-catenin in mixed cultures of n2- and [n2+Ecad]-293 (left) and n2- and [n3+Ecad]-293 cells (right). The top figures show the cellular pattern of the mixed culture (green, n2-293; red, [n2+Ecad]-293 or [n3-Ecad]-293) and immunostaining for β-catenin (white). A representative image of nine independent experiments is shown. Yellow lines indicate the positions of the densitometric tracing in the lower graphs. The graphs indicate the fluorescence intensity of β-catenin corresponding to three different homotypic or heterotypic junctions marked with the same number in the upper figures. (E) Statistical analysis for the normalized fluorescence intensity of β-catenin in mixed cultures of n2- and [n2+Ecad]-293 (left) and n2- and [n3+Ecad]-293 cells (right). Results shown are the mean ± SD; *, P < 0.001. n = 9.](https://cdn.rupress.org/rup/content_public/journal/jcb/212/5/10.1083_jcb.201509020/6/jcb_201509020_fig8.jpeg?Expires=1773841892&Signature=b48cHfc5yZ2G0F5-shlGM2OWxG8Z~0OODTXX3ZBhR~wmzPhnmSHOVyA85EeWy7mszRHkEGDMp6AddAalqNBa1ZhvCflKdfTWhp9I1wa9-Y5DCaOFKJ7M4~kRaz83SfbnRHPBXRyd-t18s-7jZoE4kj2gGeBzUQZJcv4zJ-9~-Z75w4pDOXD-LFrBCihqUnE-NArt4my4g9GHmwaRUJRVJYEp3DxJelq9g3tZSgYmBQQrUbjd4dnUFedkT4UYrMrcBGr0DHy-jBeP1TflfGrSgnW7fxJnGtNMfZhS7ias7HDkxBE7i6YUHOJf10aDw74NafGp6~S5ISgoQBSJAFbDlA__&Key-Pair-Id=APKAIE5G5CRDK6RD3PGA)
Synergistic effects of combinatorial expression of nectins and cadherins on asymmetric mosaic cellular patterning. (A) The mosaic-forming assay using neuro-2a (n2a) cells expressing different combinations of cadherins and nectins. Boundaries between the combinations of n2- and n2-n2a cells (left), n2- and n3-n2a cells (left middle), [n2+Ecad]- and [n2+Ecad]-n2a cells (right middle), and [n2+Ecad]- and [n3+Ecad]-n2a cells (right) are indicated. n2-n2a, neuro-2a nectin-2 transfectants; n3-n2a, nectin-3 transfectants; [n2+Ecad]-n2a, nectin-2 and E-cadherin transfectants; [n3+Ecad]-n2a, nectin-3 and E-cadherin transfectants. The graph shows the number of singly isolated cells in the mixtures of the two cell populations in each field. The results shown are the mean ± SD; *, P < 0.001; n.s., not significant. n = 9. (B) The mosaic-forming assay of HEK293 (293) cells expressing different cadherins and nectins. Boundaries between the combinations of n2- and n2-293 cells (top), n2- and n3-293 cells (middle), and n2- and [n3+Ecad]-293 cells (bottom) are indicated. White dotted lines indicate the center of the field. n2-293, 293 nectin-2 transfectants; n3-293, nectin-3 transfectants; [n2+Ecad]-293, nectin-2 and E-cadherin transfectants; [n3+Ecad]-293, nectin-3 and E-cadherin transfectants. (C) The graph indicates the number of singly isolated cells in the counter colony in each field. The results shown are the mean ± SD; *, P < 0.05. n = 9. (D) Distribution patterns of β-catenin in mixed cultures of n2- and [n2+Ecad]-293 (left) and n2- and [n3+Ecad]-293 cells (right). The top figures show the cellular pattern of the mixed culture (green, n2-293; red, [n2+Ecad]-293 or [n3-Ecad]-293) and immunostaining for β-catenin (white). A representative image of nine independent experiments is shown. Yellow lines indicate the positions of the densitometric tracing in the lower graphs. The graphs indicate the fluorescence intensity of β-catenin corresponding to three different homotypic or heterotypic junctions marked with the same number in the upper figures. (E) Statistical analysis for the normalized fluorescence intensity of β-catenin in mixed cultures of n2- and [n2+Ecad]-293 (left) and n2- and [n3+Ecad]-293 cells (right). Results shown are the mean ± SD; *, P < 0.001. n = 9.
