Figure S2.
The impact of physiological (5%) and atmospheric (21%) oxygen tension on cell behavior in vitro.(A) Analysis of migration in E1A;HrasG12V MEFs in 21% O2. Data are mean fold change in migrating cells ± SD, expressed relative to counterpart sgNTC control cell line. n = 3 cell lines/sgRNA; ns, not significant (P > 0.05), Mann–Whitney unpaired t test. (B) Cell viability analysis by AnnexinV/PI staining of sgNTC and sgp53(2) E1A;HrasG12V MEFs in 5% or 21% O2 cotreated with 0.2 µg/ml doxorubicin or 0.1% FBS for 24 h. n = 3 independent experiments for each sgRNA. Data are presented as mean ± SD; ****, P < 0.0001; ns, not significant (P > 0.05), two-way ANOVA, Sidak’s multiple comparison posttest. (C) Proliferation of E1A;HrasG12V MEFs in 5% or 21% O2 measured by cell counting starting 24 h after plating. n = 3 cell lines/sgRNA. Data are mean fold change in cell number ± SD; **, P < 0.005; not significant (P > 0.05), linear regression analysis. (D) Representative soft agar assays measuring anchorage-independent growth of sgNTC and sgp53(2)-targeted E1A;HrasG12V MEFs in 5% or 21% O2. Scale bar, 3.5 mm. (E) Average colony number ± SD of sgNTC and sgp53(2) E1A;HrasG12V MEFs in 5% and 21% O2. n = 3 cell lines/sgRNA, in triplicate; *, P < 0.05; ****, P < 0.0001; ns, not significant (P > 0.05), one-way ANOVA, Tukey’s multiple comparison posttest.

The impact of physiological (5%) and atmospheric (21%) oxygen tension on cell behavior in vitro. (A) Analysis of migration in E1A;HrasG12V MEFs in 21% O2. Data are mean fold change in migrating cells ± SD, expressed relative to counterpart sgNTC control cell line. n = 3 cell lines/sgRNA; ns, not significant (P > 0.05), Mann–Whitney unpaired t test. (B) Cell viability analysis by AnnexinV/PI staining of sgNTC and sgp53(2) E1A;HrasG12V MEFs in 5% or 21% O2 cotreated with 0.2 µg/ml doxorubicin or 0.1% FBS for 24 h. n = 3 independent experiments for each sgRNA. Data are presented as mean ± SD; ****, P < 0.0001; ns, not significant (P > 0.05), two-way ANOVA, Sidak’s multiple comparison posttest. (C) Proliferation of E1A;HrasG12V MEFs in 5% or 21% O2 measured by cell counting starting 24 h after plating. n = 3 cell lines/sgRNA. Data are mean fold change in cell number ± SD; **, P < 0.005; not significant (P > 0.05), linear regression analysis. (D) Representative soft agar assays measuring anchorage-independent growth of sgNTC and sgp53(2)-targeted E1A;HrasG12V MEFs in 5% or 21% O2. Scale bar, 3.5 mm. (E) Average colony number ± SD of sgNTC and sgp53(2) E1A;HrasG12V MEFs in 5% and 21% O2. n = 3 cell lines/sgRNA, in triplicate; *, P < 0.05; ****, P < 0.0001; ns, not significant (P > 0.05), one-way ANOVA, Tukey’s multiple comparison posttest.

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