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. 2020 Oct 12;11:5126. doi: 10.1038/s41467-020-18919-2

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a–c Heatmap of changes in gene expression of the indicated genes in: a f-HDF strains plus/minus CSL or NOTCH1 silencing by two different siRNAs versus siRNA controls for 3 days; b f-HDF strains infected with a lentivirus for doxycycline-inducible expression of ICN1 versus empty vector control, 5 days after ICN1 induction or cultured on JAGGED 1-coated versus IgG coated control dishes for 3 days; c CAF strains infected with two different NOTCH1 silencing lentiviruses versus empty vector control for 5 days or treated with the γ-Secretase Inhibitor DBZ (10 μM) versus DMSO vehicle alone for 5 days. Results are shown as gene expression folds (down- or up-regulation in blue and red, respectively). Individual bar plots and statistical quantification of the results are shown in Supplementary Fig. 2a–c. d Comet assays of f-HDF strains with or without CSL and NOTCH1 silencing. The percentage DNA in tail for individual cells is indicated by scatter dot plot. >184 cells were analyzed per sample. One-way ANOVA followed by Dunnett’s test. n(f-HDF strain) = 3. e Immunofluorescence analysis with antibodies against γ-H2AX of f-HDF strains plus/minus CSL and NOTCH1 silencing. >201 cells were analyzed per sample. One-way ANOVA followed by Dunnett’s test. n(f-HDF strain) = 3. n(CAF strain) = 3. f–i EdU assays of: f f-HDF strains plus/minus CSL and NOTCH1 silencing. n(cells) >201, One-way ANOVA followed by Dunnett’s test n(f-HDF strain) = 3; g m-HDF and CAF strains plus/minus siRNA mediated NOTCH1 gene silencing and plus/minus UVA irradiation. 24 h after siRNA transfection cells were mock treated or irradiated with UVA (500 mJ/cm²), followed by EdU labeling 48 h later, n(cells)> 99, two-way ANOVA between groups and two-tailed unpaired t-test within each group, n(m-HDF strain) = 3, n(CAF strain) = 3; h f-HDF, m-HDF, and CAF strains 72 h after UVA irradiation (500 mJ/cm²) versus mock treated, n(cells)>89, two-tailed unpaired t-test, n(CAF strain) = 3. n(f-HDF strain) = 3, n(m-HDF strain) = 3, and n(CAF strain) = 3; i CAF strains plus/minus shRNA-mediated NOTCH1 silencing for 7 days and plus/minus UVA irradiation as in h. n(cells) >103, two-tailed unpaired t-test, n(CAF strain) = 3. All data represented as Mean ± s.d, *p < 0.05, **p < 0.01, ***p < 0.001, ****p < 0.0001.