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. 2019 Sep 19;179(1):219–235.e21. doi: 10.1016/j.cell.2019.08.032

Figure S2.

Figure S2

In Vitro and In Vivo Assessment of UVB-Treated Cells, Related to Figure 2 and Table S2

A) western blot for p53 and GAPDH in B16F10.9 and B2905 cells 24h post irradiation in various UVB dosages. B) Immunofluorescence stains for UVB-induced cyclobutane pyrimidine dimer (CPD) in parental B2905 cells -untreated (left) and UVB irradiated (right). The UVB irradiated cells were fixed, washed and subjected to immunostaining two minutes after irradiation. Scale bar represents 200 μM (for 10x magnification) and 100 μM (for 20x magnification). C) Distribution of somatic alterations in the parental cell line, in comparison to those occurring following UVB exposure show increase in C > T alterations (distribution of added mutations is shown relative to parental). p value was calculated based on the Chi-square test D) Mutation signatures identified by DeconstructSigs for the mutation changes following UVB irradiation (UVB B2905 versus Parental cell line). E-F) In vitro proliferation of parental versus UVB irradiated B2905, and B16F10.9, respectively, starting from 500 cells at day 0. Data are mean ± SEM. G) In vivo tumor growth in mice inoculated with parental B16F10.9 (left panel) and UVB irradiated B16F10.9 (right panel). n = 5, data are representative of two independent experiments. H) Day of tumor onset for the experiment shown in G. I) In vivo tumor growth in mice inoculated with parental B2905 (Black) or UVB irradiated B2905 (red) lines, treated with anti-PD-1 or IgG control antibodies at days 6, 9, and 12 post cells inoculation (n = 11-12). Data are mean ± SEM. Comparisons between parental B2905 tumors treated with IgG or anti-PD-1 treated are depicted by asterisks, whereas comprisons between UVB B2905 tumors treated with IgG or anti-PD-1 are depcited by cross. ∗,+p < 0.05, ∗∗p < 0.01, ∗∗∗p < 0.001, one-way ANOVA followed by Tukey's post hoc test, J) Mutation signature 7, associated with UVB exposure, is identified in SCC 1 and 2 (deconstructSigs tool). K) Macroscopic tumor growth for tumors derived from UVB, SCC 1 and SCC 2 at day 15.