Figure 3. Adherens junctions regulate keratinocyte patterning.

(A, B) Immunofluorescent images of keratinocytes on day 4. Nuclei are stained with DAPI and Hoechst 33342 (blue). (A, B) Cell–cell adhesions are visualized with E-cadherin (A) and α18 labeling (B) (green). Scale bar: 50 μm. (C) Schematic diagram of experiments investigating patterns under high-calcium (1.8 mM) and low-calcium (0.06 mM) conditions. (D) Immunofluorescent images of keratinocytes under high- and low-calcium conditions. Nuclei are labeled with Hoechst 33342. Scale bar: 500 μm. (E) Pattern index in high- and low-calcium conditions. N = 4 for each group. (F) Schematic diagram of experiments investigating patterns with or without the E-cadherin-blocking antibody. (G) Immunofluorescent images of keratinocytes with or without the E-cadherin-blocking antibody. Nuclei are labeled with Hoechst 33342. Scale bar: 500 μm. (H) Pattern index with or without the E-cadherin-blocking antibody. N = 4 for each group. (I) Schematic diagram of experiments investigating patterns with or without blebbistatin, a non-muscle myosin II inhibitor. (J) Immunofluorescent images of keratinocytes with or without blebbistatin. Nuclei are labeled with Hoechst 33342. Scale bar: 500 μm. (K) Pattern index with or without blebbistatin. N = 8 for each group. (L) Immunofluorescent images of WT and CTNNA1-knockout (KO) keratinocytes. Nuclei are labeled with Hoechst 33342. Scale bar: 500 μm. (M) Pattern index of WT and CTNNA1-KO keratinocytes. N = 8 for each group. All data are presented as mean values. Data for (E, H, K) were analyzed with two-tailed Mann–Whitney U tests. Data for (M) were analyzed with the Kruskal–Wallis test followed by Dunn’s multiple comparison test. ns, not significant. *P < 0.05; **P < 0.01; ***P < 0.001.