Extended Data Fig. 9: YAP/TAZ mechanotransduction is required to preserve NE architecture.
From: YAP/TAZ activity in stromal cells prevents ageing by controlling cGAS–STING
a, Quantification of nuclear dysmorphia (related to Fig. 5a) in young control or YAP/TAZ-depleted MAFs, and in old MAFs with or without YAP reconstitution. Data (n = 3 independent experiments) are presented as mean ± s.d.; P values are derived from one-way ANOVA with Dunnett’s multiple comparison test. b, Nuclear morphology (left panel) and quantification of nuclear dysmophia (right panel) in control (siCo) and YAP/TAZ-depleted (siY/T) WI-38 cells, as assessed by Lamin A/C immunostaining. Scale bar, 10 μm. Data (n = 3 independent experiments) are presented as mean ± s.d.; P values are derived from two-sided, unpaired t-test. c, Orthogonal projection of Lamin A/C-stained nuclei (left panel, Scale bar, 1 µm) and quantification of nuclear height (right panel; n = 70 cells per condition) in control or YAP/TAZ-depleted WI-38. Data are presented as mean ± s.d.; P values are derived from two-sided, unpaired t-test. d, Pie charts showing the relative percentage of nuclear lamina defects upon Y/T depletion in WI-38 fibroblasts. At least 200 cells per condition were analyzed. e, f Nuclear morphology (right panel) and quantification (left panels) of nuclear dysmorphia in WI-38 cells plated on stiff (40 kPa) or soft (0.5 kPa) hydrogels (e) and WI-38 cells plated on large or small microislands (f), as assessed by Lamin A/C immunostaining. Data (n = 3 independent experiments) are presented as mean ± s.d.; P values are derived from two-sided, unpaired t-test. Scale bar, 10 μm. g, Nuclear morphology (left panel, Scale bar, 10 μm) and quantification (left panel) of nuclear dysmorphia in YAP/TAZ-depleted WI-38 cells, in presence or absence of Remodelin, as assessed by Lamin A/C immunostaining. Data (n = 3 independent experiments) are shown as mean ± s.d.; P values are derived from one-way ANOVA with Dunnett’s multiple comparison test. h, Quantification of EGFP-cGAS reporter signal in YAP/TAZ-depleted (siY/T) WI-38 cells, with or without Remodelin treatment. Data (n = 3 independent experiments) are shown as mean ± s.d.; P values are derived from one-way ANOVA with Dunnett’s multiple comparison test. Related to Fig. 5b. i, j SASP marker genes expression in WI-38 cells (i, n = 2 independent experiments) and primary MAFs (j, n = 3 independent experiments) upon YAP/TAZ depletion is abolished by concomitant Remodelin (Rem) treatment, as assessed by RT-qPCR analysis. In j, treatment with the STING inhibitor H-151 (STINGi) served as control. Data are shown as mean ± s.d.; P values are derived from one-way ANOVA with Dunnett’s multiple comparison test. k, In vivo super-resolution microscopy analysis (left panel, Scale bar, 1 μm) and quantification (right panel, at least n = 130 cells per condition were analyzed) of perinuclear F-Actin in dermal fibroblasts of Control and YAP/TAZ cKO mice. Data are shown as mean ± s.d.; P values are derived from two-sided, unpaired t-test.
