Supplementary Figures:Fig. 1, 2, and 3 show H1299 cells overexpressing Set7/9wt or H297A catalytic mutant and U2-OS, U2-OS Set7/9KD, andU2-OS Set7/9 KD cells expressing ectopic Set7/9wt or its H297A mutant. These cell lines were treated with different sources of DNA damage: doxorubicin (1), gamma- (2), or UV-irradiation (3). Following the treatment with genotoxic stress cells were harvested at differenttime points, stained with Propidium Iodine and analysed by FACS for their cell cycle distribution. In all the cases the lack of Set7/9 resulted ina more pronounced fraction of cells arrested in G1/S phase. Addition of the ectopically expressed Set7/9wt or H297A mutant into shRNA-Set7/9 cells resulted in different effects: Set7/9wt, but not Set7/9mut, partially restored normal DDR (DNA Damage Response) of U2-OS cells.Overexpression of Set7/9mut in H1299 cells (p53- ) despite the presence of the endogenous Set7/9 protein, also resulted in significant G1/S arrest. Collectively, these data suggest that Set7/9 is important for cell cycle progression in response to various forms of DNA Damage (DD).

Fig. 4. The bioinformatics analysis demonstrates that the positive correlation between E2F1 and TP73 expression positively affects survival of the lung cancer patients (see description of the experiment provided for Fig. 5). Left panel shows that low expression of E2F1 (green line) in lung cancer patients correlates with better survival. The two other upper panels denote groups with strong positive correlation (middle) between E2F1 and TP73 expression and negative correlation (right). The lower panel shows the rate of survival for patients with positive (green line) and negative (red line) correlations.

Fig. 5. A breast cancer data set shows that the negative correlation between Set7/9 and TP73 is associated with a better prognosis for survival. Left panel provides statistically significant correlation (p=0.015) between low expression of E2F1 and better survival (see description of the experiment provided for Fig. 5).

Table 1. A summary of Set7/9, Cyclin E, E2F1, and TP53 expression levels in six different lung cancer cell lines and their cell cycle and apoptoticresponse to DNA Damage induced by doxorubicin.

control Set7/9wt Set7/9mut

no treatment

14 hrs doxorubicin

24 hrs doxorubicin

H1299 cells (p53-) U2-OS cells (p53+)

no treatment

24 hrs doxorubicin

control Set7/9KD+Set7/9wt

Set7/9KD+Set7/9mut

Supplementary Fig.1

U2-OS-Set7/9 KD U2-OS control

0 hrs X-rays

16 hrs X-rays

48 hrs X-rays

Supplementary Fig.2

0 hrsUV

8 hrsUV

16 hrsUV

Set7/9KD (LSL) U2-OS control

Supplementary Fig.3

Group=1: green , strong positive correlation (0.53) between E2F1 and TP73Group=2: red , no (or negative) correlation (-0.53) between E2F1 and TP73

p= 0.03

GEO dataset ID: GSE36471GEO dataset Type: lung cancer

Positive correlation between  E2F1  and  TP73 expression increases survival in lung cancer, where E2F1 is an oncogene

Supplementary Fig.4

GEO dataset ID: GSE36471GEO dataset Type: lung cancer

E2F1 p= 0.016

GEO dataset ID: GSE21653GEO dataset Type: Breast Cancer

Group=1: green , strong positive correlation (0.73) between SETD7 and TP73Group=2: red , no (or negative) correlation (-0.33) between SETD7 and TP73

p= 0.025

GEO dataset ID: GSE21653GEO dataset Type: Breast Cancer

E2F1 p= 0.015

Supplementary Fig.5

Inverse correlation between SETD7 and TP73 expression in breast cancer, where E2F1 is an oncogene, increases survival

Cell LineCell cycle

checkpoint arrest

Set7/9 Cyclin E TP53 E2F1Apoptosis

(PARP cleavage)

cont dox cont dox cont dox cont dox

p53 status

H23 p53mut S/G2 ++ +++ ++ +++ ++ ++ + ++ +

H520 p53del G1/S +/- +/- + + - - + ++ +

H522 p53mut S/G2 ++ +++ + +++ + ++ + ++ +

H1650 p53mut G1/S + +/- + +/- - - + + -

H460 p53wt G1/S +++ +++ +/- +/- + +++ + + +

H1299 p53del S/G2 ++ ++ + ++ - - + ++ -

Supplementary Data: Table 1

name