EMBO/soda-nel huggingface dataset:
F-domain valency determines outcome of signaling through the angiopoietin pathway (2021)
Figure 5

Figure 5. Icosahedral F-domain scaffold restores BBB function and enhances angiogenesis after injury.

A) Brain tissue from CCI-TBI animals injected with Evans blue (5% in saline) after Sham, Icos1ø (control), or Icos1 (Tie2 super-agonist) injection at 6, 24, 48, and 72 hours post-injury was harvested 4 days post-injury.

B) Evans blue extravasation in brain tissue (black dotted-circle indicates lesion site) was quantified by fluorescence intensity in tissue-extracts from hemispheres contra- and ipsilateral to the injury. Intensity measurement from female and male mice were normalized against body mass to report changes in BBB permeability in Icos1ø and Icos1 treated animals (p = 0.015, p <0.0001; n = 6). Scale bar is 1 mm.

C, D) CCI-TBI brain sections were stained for reactive astrocytes (GFAP, blue), microglia (Iba1, green) and serum albumin (magenta) to highlight tissue margins (dotted white line) and quantify secondary damage as a function of lesion area (E; *p = 0.04, n = 4, the mean ± SEM is plotted ). Bar, 1 mm.

E) Cumulative blood extravasation into brain tissue (white outline) was imaged by albumin immunofluorescence to establish an unbiased intensity-threshold mask (red) and quantify the area of blood serum extravasation (mm2) in Icos1ø vs. Icos1 treated animals (**p = 0.01, n = 4, the mean ± SEM is plotted). Scale bar is 500 µm.

F-H) Brain tissues stained by GFAP- (magenta) and CD31-immunofluorescence (green) were imaged by confocal microscopy to measure angiogenesis as a function of vessel diameter (G; **p = 0.003, n = 4, the boxes show 25th to 75th percentiles and whiskers represent the min and max values.) and quantify the number of CD31-vessels (H; **p = 0.007, n = 4, the mean ± SEM is plotted) at the site of injury in Icos1ø and Icos1 treated animals 4 days post-injury. Scale bars in G and H is 100 µm.

I) Activation of pAKT (Magenta) was examined in Icos1ø and Icos1 treated-animals within or around vascular (CD31+, green) and pericytes (NG2+ cells, yellow) of the BBB. High-resolution confocal images (insets) reveal the prevalence of activated pAKT (magenta) co-stain (cyan arrows) in pericytes (yellow) of the BBB along vasculature (green, or dotted outline) versus reactive neuroglia (NG2+pAKT-) in the parenchyma (red arrows) in Icos1ø vs. Icos1 treated animals. Scale bar is 10 µm.

Data information: One-way ANOVA with Dunnett's test was used for multiple comparisons and a t-test was used to compare two groups.
Inhibition of the NLRP3 inflammasome improves lifespan in animal murine model of Hutchinson-Gilford Progeria (2021)
Figure 1

Figure 1. NLRP3 signalling is associated to HGPS.

(A) Western blot analysis with representative blot including lamin A/C, NLRP3, caspase 1, and actin levels in skin fibroblats from patients with HGPS, n = 2 controls and 2 patients. Positive control correspond to THP1 cells stimulated with LPS+ATP.

(B) Immunofluorescence (IF) visualization of NLRP3 (green) and nuclei (blue) in skin fibroblasts from a representative patient and control. Scale bar: 30µm

(C) Protein expression of NLRP3 and caspase 1 in lymphoblasts from control and one patient after stimulation with uric acid and cholesterol crystal.

(D) IL-1β and IL-18 medium release from lymphoblasts which were assessed after a 24 hr incubation with uric acid and cholesterol. ***P < 0.001, **P < 0.005, * treatment vs no treatment; aaaP < 0.001; aaP < 0.01 control cells vs patient cells.

(E) Western blot analysis with representative blot including NLRP3, caspase 1, IL-1β and actin levels in heart and liver tissues from wild-type and Zmpste24-/- mice.

(F) NLRP3 and Caspase 1 transcript expression levels were determined in heart and liver tissues by real-time quantitative RT-PCR. n=5 for Zmpste24+/+ and n=5 for Zmpste24-/- groups respectively. ***P < 0.001, **P < 0.005, wild-type vs Zmpste24-/- mice. Data are showed means ± SD, n = 4 mice per group.

Data information: Results are presented as the mean ± SD of three independent experiments. One-way ANOVA test was used for statistical analysis
Inhibition of the NLRP3 inflammasome improves lifespan in animal murine model of Hutchinson-Gilford Progeria (2021)
Figure 2

Figure 2. NLRP3 inhibition by MCC950 ameliorates skin fibroblasts and Zmpste24-deficient mice progeroid phenotypes.

(A) Cell growthn (lower subpanel left patient 1 and right patient 2) and morphological aspect (upper subpanel) with MCC950 determined in healthy and representative HGPS fibroblasts. **P < 0.01, *P < 0.05 patient cells no treatment vs treatment. Results are presented as the mean ± SD of three independent experiments.

(B) Western blot analysis with representative blots including lamin A/C, NLRP3, IL-1β and actin levels in skin fibroblats from control and HGPS patient after 48 hr of vehicle and MCC950 treatment.

(C-E) Representative fluorescence images of HGPS and control fibroblasts to evaluate the effect of the MCC950 in the nuclear morphology (D) and NLRP3 expression (E). Scale bar: 30µm. Results are presented as the mean ± SD of three independent experiments. ***P < 0.01 patient vs control cells, aaaP < 0.01 patient cells no treatment vs treatment. One-way ANOVA test was used for statistical analysis.

(F) Kaplan-Meier graph showing a significant increase in the maximum lifespan in WT mice compared with Zmpste24-/- mice. N=7 per group.

(G) Body weights evolution of the groups over time. Data are showed means ± SD, n = 6 mice per group.

(H) Analysis of serum concentrations of IL-1β measured by ELISA. N=6 per group. Data are shown as means ± SD. ***P < 0.001, wild-type vs Zmpste24-/- mice; aaaP < 0.001; vehicle vs MCC950.
NBS1 promotes the endonuclease activity of the MRE11-RAD50 complex by sensing CtIP phosphorylation (2019)
Figure 1

Figure 1 - NBS1 in trans with MR and pCtIP cleaves DNA similarly as MRN-pCtIP.

A representative nuclease assay with MR, MBP-NBS1-his (denoted MBP-NBS1), MRN and pCtIP on 5'-end labeled 70-bp dsDNA with all ends blocked with streptavidin. Samples were separated on 15 % denaturing polyacrylamide gel.

Quantitation of nuclease assays such as in (A). Averages shown; n ≥ 3; error bars, SEM.
NBS1 promotes the endonuclease activity of the MRE11-RAD50 complex by sensing CtIP phosphorylation (2019)
Figure 2

Figure 2 - FHA and BRCT domains of NBS1 are important for MR-pCtIP stimulation while MRE11-NBS1 interaction is essential.

A schematic representation of purified recombinant wild type NBS1 and variants. All constructs were MBP tagged at the N- and his-tagged at the C-terminus.

The effect of mutations in FHA and/or BRCT domains of NBS1 on the nuclease of MR and pCtIP. Quantitation of experiments such as shown in Fig EV2D. Averages shown; n ≥ 4, error bars, SEM. Statistical significance denotations represent the analysis between wild type full-length NBS1 (in black) and the corresponding concentrations of the NBS1 variants; ns (p > 0.05, not significant), * (p < 0.05), ** (p < 0.01), *** (p < 0.001), two-tailed t-test.

Analysis of pCtIP binding to NBS1 variants. Anti CtIP antibody was immobilized on Protein G agarose, bound to pCtIP, and tested for interactions with the indicated NBS1 variants (see cartoon at the top). Western blot of eluates (right part) was performed with anti-CtIP and anti-NBS1 antibodies. Left part (input) indicates that the anti-NBS1 antibody recognizes the NBS1 constructs to a similar level.

Analysis of phosphorylated CtIP (pCtIP, mock-treated) or λ phosphatase treated CtIP (λCtIP) interactions with NBS1. The CtIP variants were immobilized on protein G agarose, and incubated with NBS1 (see cartoon on the left). The western blot was performed with anti CtIP and anti NBS1 antibodies.

The effect of the MRE11 interaction region (MIR) of NBS1 on the nuclease of MR and pCtIP. Quantitation of experiments such as shown in Fig EV2E. Averages shown, n ≥ 3, error bars, SEM. The quantitation of wild type NBS1 (NBS1 WT, in black) is the same as in panel (B) and is shown again for reference. Statistical significance denotations represent the analysis between wild type full-length NBS1 (in black) and the corresponding concentrations of the NBS1 variants; ns (p > 0.05, not significant), *** (p < 0.001), two-tailed t-test.
NBS1 promotes the endonuclease activity of the MRE11-RAD50 complex by sensing CtIP phosphorylation (2019)
Figure 3

Figure 3 - Without pCtIP, NBS1 promotes the MR endonuclease independently of its FHA and BRCT domains.

Nuclease assays with MR and various NBS1 fragments (50 nM) on oligonucleotide-based 5'-end labeled dsDNA. The reaction buffer contained 60 mM NaCl and reactions were incubated for 2 h.

Quantitation of experiments such as shown in (A). Averages shown; n ≥ 3, error bars, SEM.
NBS1 promotes the endonuclease activity of the MRE11-RAD50 complex by sensing CtIP phosphorylation (2019)
Figure 4

Figure 4 - The function of NBS1 in DNA end resection in vivo.

A schematic representation of the NBS1 knockdown-rescue experiments in U2OS cells.

In vivo resection assay using BrdU staining under native conditions to directly detect ssDNA at sites of DSBs. Shown are representative images.

In vivo resection assay using RPA2 staining as a marker for ssDNA present at sites of DSBs. Shown are representative images.

Quantitation of the experiment shown in (B). At least 100 cells were assessed per condition. The horizontal line represents a median, the box spans from 25th to 75th percentile and the whiskers from 10th to 90th percentile.

Quantitation of the experiment shown in (C). At least 100 cells were assessed per condition. The horizontal line represents a median, the box spans from 25th to 75th percentile and the whiskers from 10th to 90th percentile.

Quantitation of the single strand annealing (SSA) reporter assay in U2OS cells. All cells were treated with DMSO; where indicated, both PFM03 (50 μM) and PFM39 (50 μM) MRE11 inhibitors were added. Averages shown; n = 4, error bars, SEM. Insert, western blot analysis of NBS1 and CtIP protein levels upon depletion with siRNA.

Quantitation of resection activity in NBS1∆N cells complemented with wild type (WT) or NBS1 R28A K160M mutant and either mock depleted (siCtrl) or depleted of CtIP (siCtIP). At least 300 cells were assessed per condition. The horizontal line represents a median, the box spans from 25th to 75th percentile and the whiskers from 10th to 90th percentile. Statistical significance was determined using ordinary one-way ANOVA with Tukey's multiple comparison test, **** (p<0.0001).
NBS1 promotes the endonuclease activity of the MRE11-RAD50 complex by sensing CtIP phosphorylation (2019)
Figure 5

Figure 5 - Phosphorylation of CtIP is partially dispensable for MR stimulation in the absence of NBS1.

Nuclease assays with MR and pCtIP on 3'-end labeled dsDNA, incubated for 2 h.

Quantitation of experiments such as shown in (A). Averages shown; n = 2; error bars, range.

Nuclease assays with MRN, MR and various concentrations of pCtIP (mock-treated) or λCtIP on 5'-end labeled dsDNA, incubated for 2 h.

Quantitation of experiments such as shown in (C). Averages shown; n = 3; error bars, SEM.

Phosphorylated CtIP was mock-incubated (pCtIP, lane 2) or treated with λ phosphatase (λCtIP, lane 3). The proteins were separated on a 10% polyacrylamide gel and stained with Coomassie brilliant blue.

Quantitation of kinetic nuclease assays with MRN and pCtIP (mock-treated) or λCtIP. Averages shown; n = 3; error bars, SEM. Statistical denotations, ** (p < 0.01), two-tailed t-test.

Quantitation of nuclease assays as in (F), but with MR complex instead of MRN. Averages shown; n = 3; error bars, SEM. Statistical denotations, ns (p > 0.05), two-tailed t-test.

Nuclease assays with MR, MRN, pCtIP or pCtIP T847A, as indicated, on 5'-end labeled dsDNA. Reactions were incubated for 2 h.

Quantitation of experiments such as shown in (H). Averages shown; n = 4; error bars, SEM.
NBS1 promotes the endonuclease activity of the MRE11-RAD50 complex by sensing CtIP phosphorylation (2019)
Figure 6

Figure 6 - CtIP phosphorylation is dispensable for its interaction with MRE11 and RAD50.

Interaction between MRE11 and CtIP. MRE11 was immobilized on antibody-coupled Protein G agarose, and incubated with pCtIP or λCtIP (see cartoon). Western blot analysis of bound proteins was performed with anti MRE11 and anti CtIP antibodies.

Interaction between RAD50 and CtIP. RAD50-FLAG was immobilized on anti-FLAG affinity resin and incubated with pCtIP and λCtIP (see cartoon). Western blot analysis of bound proteins was performed with anti FLAG and anti CtIP antibodies.

Interaction between MRE11, MR or MRN and pCtIP or λCtIP, as indicated. Phosphorylated (pCtIP) or λ phosphatase-treated CtIP (λCtIP) were immobilized on antibody-coupled Protein G agarose, and incubated with MRE11, MR or MRN complexes (see cartoon). Bound proteins were analyzed by western blot using anti CtIP and anti MRE11 antibodies.
NBS1 promotes the endonuclease activity of the MRE11-RAD50 complex by sensing CtIP phosphorylation (2019)
Figure 7

Figure 7 - NBS1 is not required for DNA cleavage opposite to a nick.

Representative nuclease assays with MRE11, MR, MRN and pCtIP, as indicated, on 5'-end labeled dsDNA with a single streptavidin-blocked end (S). The substrate contained 5 phosphorothioate bonds (PTO) at both 3'-ends to prevent exonucleolytic DNA degradation. Reactions were incubated for 2 h.

Representative nuclease assays as in (A), but with a substrate containing a nick in the top oligonucleotide (20 nt away from the 3'-end, see cartoon).

Quantitation of experiments such as shown in (A), with pCtIP. Averages shown; n ≥ 4; error bars, SEM.

Quantitation of experiments such as shown in (B), with and without pCtIP. Averages shown; n = 3; error bars, SEM.
Pharmacological CDK4/6 inhibition reveals a p53-dependent senescent state with restricted toxicity (2021)
Figure 1

Figure 1. CDK4/6i treatment induces a state of cellular senescence dependent on p53 activity.

(A-B) Human fibroblasts (BJ) were treated with vehicle (water for 8 times 24 hours) or abemaciclib (1 μM for 8 times 24 hours). The population doubling over the 8 days treatment is plotted (A). 1 or 8 dpt, cells were incubated with EdU for 20 hours and stained and quantified (n=9 samples from 3 independent experiments; scale bar, 150 μm) (B).

(C) Chromatin was extracted from BJ cells treated with vehicle or abemaciclib (1 μM for 8 times 24 hours) or nutlin-3a (10 μM for 8 times 24 hours) or doxorubicin (250 nM for 24 hours) and ChIP assays using an antibody against p53 were performed. qRT-PCR was performed using primers amplifying promoter regions of CDKN1A, GADD45A and MDM2 genes containing p53 binding sites. Values indicate fold enrichment relative to the vehicle group (n=3 independent experiments).

(D) BJ cells transduced with a p53 reporter were treated with nutlin-3a (10 μM, positive control) or abemaciclib (1 μM for 1 or 4 or 8 times 24 hours) and luciferase activity measured after treatments at the indicated time points (n=3 independent experiments).

(E) 3*103 BJ cells of the indicated genotypes were re-plated after vehicle or abemaciclib treatment (1 μM) and stained with 0.2% crystal violet 8 dpt (n=3 independent experiments).

(F) 8 dpt, treated BJ cells were incubated with EdU for 20 hours and stained (n=9 samples from 3 independent experiments).

(G) 3*103 mouse dermal fibroblasts (MDFs) of the indicated genotypes were re-plated after vehicle or abemaciclib treatment (4 μM for 8 times 24 hours) and stained with 0.2% crystal violet 8 dpt (n=3 independent experiments).

(H) 8 dpt, treated MDFs were incubated with EdU for 20 hours and stained (n=9 samples from 3 independent experiments).

(I) RNA was isolated from vehicle or abemaciclib (1 μM) treated scramble/shp53 BJ cells and mRNA for the indicated genes quantified by qRT-PCR relative to tubulin (n=3 independent experiments).

Data information: Data are means ±SD. Unpaired student t-test (A and H). One-way ANOVA (B, D and F). Two-way ANOVA (C and I). *p<0.05, **p<0.01, ***p<0.001. dpt, days post treatment.
Pharmacological CDK4/6 inhibition reveals a p53-dependent senescent state with restricted toxicity (2021)
Figure 2

Figure 2. CDK4/6i promotes premature senescent cells in vivo without toxicity.

(A-E) p16-3MR mice were treated with vehicle (PBS, 7 consecutive days), doxorubicin (5 mg/kg, 3 consecutive days) or abemaciclib (50 mg/kg, 7 consecutive days). n=6 mice/group. 14 dpt, bioluminescence was visualized and quantified by the IVIS spectrum in vivo imaging system, as shown by representative bioluminescence images (A) and quantification (B). RNA isolated from kidneys and mRNA encoding p16 quantified by qRT-PCR (C). Representative images (D) to visualize SA-β-gal activities in vehicle-, doxorubicin- or abemaciclib-treated mouse kidney sections at 15 dpt (arrows indicated positive area; scale bar, 1 mm; n=3 mice/group) and quantified (E).

(F) p16-3MR mice were treated with abemaciclib (50 mg/kg, 7 consecutive days) ± pifithrin-α (2 mg/kg, 7 consecutive days). Values indicate the ratio of post-treatment bioluminescence to pre-treatment (n=5 mice/group).

(G) RNA was isolated from abemaciclib ± pifithrin-α treated kidneys and mRNA encoding p16 and p21 genes was quantified by qRT-PCR and normalized on tubulin (n=5 mice/group).

(H-N) For the doxorubicin or abemaciclib treated mice (n=6 mice/group), physical performance was measured by rotarods assay at 15 dpt (H), grip strength meter at 7 dpt and 14 dpt (I), and hanging tests were performed at 7 dpt and 14 dpt and normalized to weights (J). (K) Relative weight changes were calculated at 7 dpt and 14 dpt. Red blood cells (L) and white blood cells (M) were counted at 15 dpt. Percentage of T cells, B cells, granulocytes and macrophages were determined by flow cytometry analysis (N).

Data information: Data are means ±SD. One-way ANOVA (B, C, E, H, L and M). Unpaired student t-test. (F). Two-way ANOVA (G, I, J, K and N). *p<0.05, **p<0.01, ***p<0.001, N.S.=not significant. dpt, days post treatment.
Pharmacological CDK4/6 inhibition reveals a p53-dependent senescent state with restricted toxicity (2021)
Figure 3

Figure 3. CDK4/6i-induced senescence lacks NF-κB activity and NF-κB-driven SASP components.

(A) 8 dpt, cells were stained for γ-H2AX (scale bar, 60 μm) and quantified (vehicle=288 cells, doxorubicin=280 cells, palbociclib=276 cells, abemaciclib=266 cells from 3 independent experiments).

(B) BJ cells transduced with a NF-κB reporter were treated with vehicle, doxorubicin, paclitaxel (50 nM for 24 hours), palbociclib (1 μM for 8 times 24 hours) or abemaciclib and luciferase activity measured 8 dpt. TNFα treatment was used as positive control (n=3 independent experiments).

(C and D) Venn plot of RNA-sequencing datasets of treated cells at 8 dpt (n=3 independent samples, sequenced together) (C). Heatmap of the SASP genes (D) (EV Table 2) for doxorubicin- or abemaciclib-treated groups (8 dpt) relative to vehicle-treated group from the RNA-sequencing datasets (p-value<0.01 was regarded as significant).

(E) 8 dpt, qRT-PCR of indicated NF-κB associated SASP genes was performed using indicated treated BJ cells 8 dpt (n=3 independent experiments).

(F) Serum-free conditioned media (CM) was collected from vehicle-, doxorubicin-, paclitaxel-, palbociclib- or abemaciclib-treated BJ cells 8 dpt and expression levels of CXCL1 and CCL5 measured by ELISA (n=6 independent experiments).

(G) Proteins in the serum-free conditioned media (CM) collected from doxorubicin- or abemaciclib-treated BJ fibroblasts (8 dpt) were measured by cytokine array. Purple boxes indicated internal positive controls from each membrane; Red boxes highlighted the most typical SASP factors.

(H-K) p16-3MR mice were treated with vehicle (PBS, 7 consecutive days), doxorubicin (5 mg/kg, 3 consecutive days) or abemaciclib (50 mg/kg, 7 consecutive days). 15 dpt, plasma was collected and expression levels of CXCL1 measured by ELISA (H), protein lysate obtained from drug treated kidneys to quantify IL6 by ELISA (I), RNA isolated from kidneys and mRNA encoding indicated NF-κB associated genes (J) quantified by qRT-PCR. n=6 mice/group. From a subset of the mice, RFP+ cells were sorted from the renal cortex using FACS and mRNA encoding indicated NF-κB associated pro-inflammatory genes quantified by qRT-PCR. The values of fold change over vehicle were plotted in the heatmap. n=4 mice/group (K).

(L) Cell-free plasma was derived from breast cancer patients treated with paclitaxel (n=10) or palbociclib (n=9) and the expression levels of CXCL1, CCL2, CCL5 and MMP1 quantified by ELISA. The values represent the ratio between on-treatment and pre-treatment levels for each patient.

Data information: Data are means ±SD. One-way ANOVA (A, B, F, H and I). Two-way ANOVA (E and J). Unpaired student t-test (L). *p<0.05, **p<0.01, ***p<0.001, N.S.= Not significant. Doxo, doxorubicin. Abe, abemaciclib. dpt, days post treatment.
Pharmacological CDK4/6 inhibition reveals a p53-dependent senescent state with restricted toxicity (2021)
Figure 4

Figure 4. p53-associated SASP factors are induced during CDK4/6i-induced senescence.

(A) Human fibroblasts (BJ) were treated with vehicle (water for 8 times 24 hours), abemaciclib (1 μM for 8 times 24 hours) or doxorubicin (250 nM for 24 hours). RNA-sequencing was performed with treated cells at 8 dpt (n=3 independent samples, sequenced together) and heatmap of the p53-associated SASP for doxorubicin- or abemaciclib-treated groups (8 dpt) relative to vehicle-treated group from the RNA-sequencing datasets (p-value<0.01 was regarded as significant).

(B) Human BJ fibroblasts, hTERT-RPE1 epithelial cells and lung mesenchymal stem cells were treated with vehicle (DMSO for 8 times 24 hours) or doxorubicin (250 nM for 24 hours) or paclitaxel (50 nM for 24 hours) or palbociclib (1 μM for 8 times 24 hours) or abemaciclib (1 μM for 8 times 24 hours). At 8 dpt, RNA was isolated and mRNA for the indicated p53-associated SASP genes quantified by qRT-PCR relative to tubulin (n=3 independent experiments).

(C) Chromatin was extracted from BJ fibroblasts treated with vehicle or abemaciclib or nutlin-3a and ChIP assays using an antibody against p53 were performed. qRT-PCR was performed using primers amplifying the promoter region of IGFBP3 containing p53 binding sites. Values indicate fold enrichment relative to the vehicle group (n=4 independent experiments).

(D and E) RNA was isolated from vehicle, abemaciclib (1 μM) (D) or palbociclib (1 μM) (E) treated scramble/shp53 BJ cells and quantified by qRT-PCR for IGFBP3 or LIF genes (n=3 independent experiments).

(F) RNA isolated from kidneys treated with doxorubicin or abemaciclib and mRNA encoding indicated p53-associated SASP genes quantified by qRT-PCR (n=6 mice/group).

(G) RNA was isolated from kidneys of p16-3MR mice treated with abemaciclib (50 mg/kg, 7 consecutive days) ± pifithrin-α (2 mg/kg, 7 consecutive days), and mRNA encoding p53 associated SASP genes was quantified by qRT-PCR and normalized on tubulin (n=5 mice/group).

(H) Cell-free plasma was derived from breast cancer patients treated with paclitaxel (n=10) or palbociclib (n=9) and the expression levels of IGFBP3 quantified by ELISA. The values represent the ratio between on-treatment and pre-treatment levels for each patient.

(I) RNA was isolated from vehicle or nutlin-3a (1 μM for 8 times 24 hours) treated BJ cells at 8 dpt and quantified by qRT-PCR for p53 target cell cycle genes, SASP genes (n=3 independent experiments).

(J) Mouse dermal fibroblasts (MDF) or MDF-p53-/- fibroblasts were transfected with empty vector (EV) or p53-overexpressing vector (p53OE), then the cells were treated with vehicle (water) or doxorubicin (250 nM for 24 hours) and qRT-PCR was performed for Igfbp3 and Lif genes (n=3 independent experiments).

Data information: Data are means ±SD. Two-way ANOVA (B, D, E, F, G, I and J). One-way ANOVA (C). Unpaired student t-test (H). *p<0.05, **p<0.01, ***p<0.001. EV, empty vector, dpt, days post treatment.