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LEGENDS TO SUPPLEMENTARY FIGURES
Suppl. Fig. 1. (A) Quantification of p53 and p21 protein levels determined by western blot in
Fig.1A using the ImageJ software, normalized to γ-tubulin and presented as fold change relative to
the level of p53 and p21 in RPE1 and HCT116 at the last day of irradiation (IR) and after 120h of
recovery (REC). Results are presented as mean±SD (n=3, ANOVA, *P<0.05 vs T0). (B)
Quantification of mitochondrial protein levels determined by western blot in Fig.1C using the
ImageJ software, normalized to γ-tubulin and presented as fold change. Results are presented as
means ± SD (n=3, ANOVA, *P<0.05 vs T0).
Suppl. Fig. 2. Relative mRNA quantification of PGC-1β target genes POLG, MFN1, MFN2,
COX5B, UQCRQ and NDUFB5 in RPE1 and HCT116 upon IR. Data are mean±SD (n=3, Paired ttest, *P<0.05 vs T0).
Suppl. Fig. 3. FACS analysis showing the fluorescence shift of MitoTracker Red (MTR) upon
irradiation of RPE1 (A), HCT116 (B) and HPS11 (C) cells. For each panel, the left hand side bar
graph displays the quantification of MTR fluorescence as mean±SD (n=3, t-test, *P<0.001 vs T0),
whereas the right hand side bar graph displays quantification of MTR fluorescence normalized on
Calcein-AM fluorescence as mean±SD (n=3, t-test, *P<0.001 vs T0).
Suppl. Fig. 4. Relative expression of p21 in HCT116p53 before (T0) and after irradiation (IR) was
evaluated as a control for p53 activation. Data are mean±SD (n=3, t-test, *P<0.05 vs T0).
Suppl. Fig. 5. Viability of (A) RPE1, (B) HCT116, (C) HPS11 and (D) HCT116TP53KO cells at the
last day of radiation treatment (IR) and after 120h of recovery (REC). Data are mean±SD (n=4, ttest, *P <0.05 vs T0; #P<0.05 vs IR).
Suppl. Fig. 6. Quantification of mitochondrial protein levels determined by western blot in Fig.3F
using the ImageJ software, normalized to γ-tubulin and presented as fold change relative to the
levels of mitochondrial protein in RPE1 and HCT116 at the last day of irradiation (IR) and after
120h of recovery (REC). Results are presented as means±SD (n=3, t-test, *P<0.05 vs T0).
Suppl. Fig. 7. (A) Quantification of HIF1α levels determined by western blot in Fig.4A using the
ImageJ software, normalized to γ-tubulin and presented as fold change relative to the level HIF1α in
HCT116 at the last day of irradiation (IR) and after 120h of recovery (REC). Results are presented
as means±SD (n=3, t-test, *P<0.05 vs T0). (B) Quantification of MDM2 levels determined by
western blot in Fig.4C using the ImageJ software, normalized to γ-tubulin and presented as fold
change relative to the level MDM2 in HCT116 at the last day of irradiation (IR) and after 120h of
recovery (REC). Results are presented as means±SD (n=3, t-test, *P<0.05 vs T0).
Suppl. Fig. 8. (A) Quantification of HIF1α levels determined by western blot in Fig.4D using the
ImageJ software, normalized to γ-tubulin, in HCT116 cells over-expressing HIF1α (HCT116HIF1α)
upon over-expression of MDM2 (+) compared to clones not over-expressing MDM2 (-). (B)
Quantification of degradation of HIF1 mutated on the three residues subjected to hydroxylation
(HIF1αTM), in presence and absence of over-expressed MDM2, determined by western blot in Fig.
4E using the ImageJ software, normalized to γ-tubulin. (C) Quantification of HIF1α levels
determined by western blot in Fig.4F using the ImageJ software, normalized to γ-tubulin, in
presence and absence of over-expressed MDM2, upon forced pharmacological HIF1 stabilization
by 1M DMOG.
Suppl. Fig. 9. (A) Western blot analysis of HIF1 in HCT116 cells before (T0) and after irradiation
(IR) in absence and in presence of 1µM DMOG. Gamma-tubulin was used as a loading control.
One representative experiment of three is shown. (B) Relative expression of BNIP3L and LDHA in
RPE1 cells before (T0) and after irradiation (IR) in absence and in presence of 1µM DMOG. Data
are mean±SD (n=3, t-test, *P<0.05).
Suppl. Fig. 10. Relative mtDNA copy number after γ-rays treatment in HCT116 cells with HIF1α
stabilized by 250nM DFO treatment. Data are mean±SD (n=3).
Suppl. Fig. 11. Relative mRNA quantification of PGC-1β target genes POLG, MFN1, MFN2,
COX5B, UQCRQ and NDUFB5 after irradiation in HCT116 cells with stabilized HIF1 after 1M
DMOG treatment. Data are mean±SD (n=3, Paired t-test, *P<0.05 vs T0).
Suppl. Fig. 12. Quantification of mitochondrial protein levels determined by western blot in Fig.4I
using the ImageJ software, normalized to γ-tubulin, in HCT116 cells with stabilized HIF1 after
1M DMOG treatment and irradiated (n=3, t-test, *P<0.05 vs T0).
Suppl. Fig. 13. (A) Relative mtDNA copy number in RPE1 cells before (T0) and after irradiation
(IR) in absence and in presence of 1µM DMOG. Data are mean±SD (n=3). (B) Western blot
analysis of mitochondrial proteins. One representative experiment of 3 is shown. (C) Relative PGC1β expression after γ-rays treatment in RPE1 cells before (T0) and after irradiation (IR) in absence
and in presence of 1µM DMOG. Data are mean±SD (n=3).
Suppl. Fig. 14. Quantification of p53, p21, MDM2 and p16 protein levels determined by western
blot in Fig.5A using the ImageJ software and normalized to γ-tubulin, in RPE1 and HCT116 cells
with stabilized HIF1 after 1M DMOG treatment at recovery after irradiation (REC). Results are
presented as mean±SD (n=3, ANOVA, *P<0.05 vs T0).
Suppl. Fig. 15. β-galactosidase staining of RPE1 and HCT116 cells irradiated and treated with
250nM DFO. One representative experiment of 3 is shown. Magnification: 10X.
Suppl. Fig. 16. Western blot analysis of MDM2 in HCT116TP53KO and HPS11 cells after irradiation.
Gamma-tubulin was used as a loading control. One representative experiment of 3 is shown.