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. 2019 Aug 1;20(9):e48084. doi: 10.15252/embr.201948084

Figure 3. Nuclear flattening is sufficient to activate c‐Jun and YAP independently of actomyosin contractility and cell spreading area.

Figure 3

  1. Representative cells stained for p‐Jun Ser63 (magenta) and for DNA (cyan) in Ctrl, Blebb, and Blebb + AP conditions. Scale bar = 10 μm. Nuclear heights were measured using Hoechst staining.
  2. Corresponding quantifications of p‐Jun Ser63 nuclear intensity. Data are presented as mean ± s.e.m. (n = 59 minimum from two independent experiments, ****P < 0.001 one‐way ANOVA—Tukey's multiple comparisons post‐test).
  3. Representative cells stained for YAP (magenta) and for DNA (cyan) in Ctrl, Blebb, and Blebb + AP conditions. Scale bar = 10 μm. Nuclear heights were measured using Hoechst staining.
  4. Corresponding quantifications of YAP activity (ratio nucleus/cytosol). Data are presented as mean ± s.e.m. (n = 82 minimum from four independent experiments, ****P < 0.001 one‐way ANOVA—Tukey's multiple comparisons post‐test).
  5. Quantifications of YAP activity (nucleo‐cytoplasmic ratio) and p‐Jun Ser63 nuclear intensity in Ctrl, Blebb, and Blebb +AC condition and in Ctrl, Y27632, and Y27632 + AP. Data are presented as mean ± s.e.m. (n = 19 minimum, ****P < 0.001 Tukey's multiple comparisons post‐test).
  6. Quantifications of YAP activity (nucleo‐cytoplasmic ratio) and p‐Jun Ser63 nuclear intensity in Ctrl, cytochalasin D (CytoD), and CytoD + AP condition. Data are presented as mean ± s.e.m. (n = 32 minimum from two independent experiments, ****P < 0.001 Tukey's multiple comparisons post‐test).
  7. Quantifications of YAP activity (nucleo‐cytoplasmic ratio) and p‐Jun Ser63 nuclear intensity in cells depleted or not for SUN1 and SUN2. Data are presented as mean ± s.e.m. (n = 28 minimum, t‐test, ****P < 0.001).
  8. Representative cells cultured on circular micropatterns with surfaces of 1,500 and 500 μm2 and stained for p‐Jun Ser63 (magenta), for YAP (cyan), and for DNA (green). Additionally, cells were cultured on the micropatterns of 500 μm2 and an agarose pad was used to flatten their nuclei. Scale bar = 10 μm. Nuclear heights were measured using Hoechst staining.
  9. Corresponding quantifications of YAP activity (nucleo‐cytoplasmic ratio) and p‐Jun Ser63 nuclear intensity. Data are presented as mean ± s.e.m. (n = 15 minimum ****P < 0.001 one‐way ANOVA—Tukey's multiple comparisons post‐test).