Fig. 7: NOTCH1 silencing or inhibition impair CAFs proliferation.

a EdU assays of CAF strains plus/minus NOTCH1 silencing (7 days) and of the same CAF strains with CRISPR-mediated disruption of the TP53 gene (as characterized in Supplementary Fig. 5a, b) plus/minus NOTCH1 silencing (7 days). >196 cells were analyzed per condition. n(CAF strain) = 3, n(CAF CRISPR TP53 strain) = 3. b Clonogenicity assays of CAF strains plus/minus shRNA-mediated NOTCH1 silencing (10 days), of CAF strains with CRISPR-mediated disruption of the TP53 gene plus/minus NOTCH1 silencing (10 days), and of three f-HDF strains plus/minus shRNA-mediated NOTCH1 silencing (10 days). n(technical replicates/condition) = 3, n(CAF strain) = 3, n(f-HDF strain) = 3, and n(CAF CRISPR TP53 strain) = 2. c EdU assays of CAF strains (#13 and 16) plus/minus CRISPR-mediated disruption of TP53 and of f-HDF strains (#5 and 6) after 5 days of treatment with DBZ (10 μM) versus DMSO vehicle alone. >125 cells were analyzed per condition. n(CAF strain) = 2, n(CAF CRISPR TP53 strain) = 2, and n(f-HDF strain) = 2. d EdU assays of CAF strains plus/minus treatment with the γ-secretase inhibitors DAPT and RO4929097 (10 μM) versus DMSO vehicle alone for 5 days. >143 cells were analyzed per condition. n(CAF strain) = 3. e EdU assays of SCC13 strain after 5 days of treatment with DBZ (10 μM) versus DMSO vehicle alone. >317 cells were analyzed per condition. n(SCC13) = 1. a–e Values for each strain are indicated as dots with mean ± s.d. two-tailed unpaired t-test, *p < 0.05. f CellTiter-Glo analysis of CAF strains infected with two NOTCH1 silencing lentiviruses versus empty vector control (for 7 days) plus/minus concomitant treatment with the ATM inhibitor KU-60019 (2 µM) at the indicated days after viral infection. Results are presented as luminescence intensity values relative to day 1. Data are shown as mean of values in triplicate dishes ±sd. Two-way ANOVA followed by Tukey’s multiple comparisons test, ****p < 0.0001. n(CAF strain) = 3.