Figure 3. Notch1 cortical signaling stabilizes adherens junctions and cortical actin organization.

(A) Super resolution by optical pixel reassignment (SoRa) fluorescence micrographs SCR, NOTCH1^KO, and ICD^KO monolayers immunostained for E-Cadherin (black). Scale bar, 10 μm. (B) Quantification of the frequency of focal adherens junctions. n ≥ 12 fields of view from three independent experiments. (C) SoRa fluorescence micrographs of SCR, NOTCH1^KO, and ICD^KO monolayers labeled with phalloidin (black). Scale bar, 10 μm. (D) Intensity of cortical actin at cell–cell junctions, quantified from phalloidin-stained micrographs. n ≥ 12 fields of view from three independent experiments. (E) Traction force microscopy traction maps averaged from ten fields of view from SCR, NOTCH1^KO, and ICD^KO monolayers. Scale bar, 20 μm. (F) Quantification of relative integrated monolayer tractions. n ≥ 11 traction force measurements from three independent experiments. (G) Fluorescence micrographs of wild-type cells treated with DMSO or 10 μM DAPT for two hours and immunostained with E-Cadherin (black). Scale bar, 10 μm. (H) Quantification of the frequency of focal adherens junctions in DMSO and DAPT treated cells. n ≥ 13 fields of view from three independent experiments. (I) Intensity of cortical actin at cell–cell junctions, quantified from phalloidin-stained micrographs of DMSO and DAPT treated cells. n ≥ 13 fields of view from three independent experiments. For plots B, D, and F, mean ± SEM; one-way ANOVA with Tukey’s post-hoc test, **p<0.01, ***p<0.001, ****p<0.0001. For plots H and I, mean ± SEM; two-tailed unpaired t test, **p<0.01, ****p<0.0001.