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Supplementary Figure 1 A B HEC1A PTEN +/+ HEC1A PTEN -/- 16 HEC1A PTEN -/- 22 PTEN RAD51 % Cells with RAD51 foci 40 -IR +IR 30 20 10 * *! TUBULIN 0 HEC1A PTEN+/+ HEC1A PTEN-/- 16 HEC1A PTEN-/- 22 C D 1.0 HR Efficiency 0.8 0.6 0.4 0.2 * * Surviving fraction 1.0 0.8 0.6 0.4 0.2 SF50 (M) HEC1A PTEN +/+ >1.0E-05 HEC1A PTEN -/- -/-22 22 1.1E-05 HEC1A PTEN -/- -/-16 16 9.4E-06 0.0 HEC1A PTEN+/+ HEC1A PTEN-/- 16 HEC1A PTEN-/- 22 0.0 0 10-9 10-8 10-7 10-6 KU0058948 (M) KU0058948 (M) 10-5

Supplementary Figure 2 "H2AX Merged HCT116 PTEN+/+ Rad51 - IR HCT116 PTEN-/- Nuclei - IR + IR + IR

Supplementary Figure 3 Surviving fraction 1.0 0.8 0.6 0.4 DU145 MCF-7 HeLa SUM159 BT474 MDA-MB-231 HCC70 PC3 RPMI-7951 SW1783 EC50 SF50 (M) 7.72E-07 4.60E-07 6.68E-07 7.94E-07 0.2 A172 UM-UC3 2.94E-08 1.71E-08 MDA-MB-468 2.66E-09 0.0 0 10-9 10-8 10-7 10-6 10-5 COLO 829 9.44E-09 KU0058948 (M)

Supplementary Figure 4 EMPTY VECTOR RAD51 cdna RAD51! Tubulin

Supplementary Table 1 Cell Line PTEN PIK3CA TP53 BRAF KRAS MLH1 MSH6 CDKN2A A172 * COLO 829 * * * HCC70 * * MDA-MB-468 * * PC3 * * RPMI-7951 * * * * SW1783 * * UM-UC3 * * * * BT474 * * HEC1A * * * HCT116 * * * * MCF7 * * SUM-159 * MDA-MB-231 * * * * DU145 * * * HeLa Cell Line Mutation PTEN PIK3CA TP53 BRAF KRAS MLH1 MSH6 CDKN2A A172 AA p.r55fs*1 CDS c.165_1212del1048 glioma Zygosity Homozygous COLO 829 AA p.? p.v600e p.a68fs*51 CDS c.493_634del142 c.1799t>a c.203_204delcg melanoma Zygosity Homozygous Heterozygous Homozygous HCC70 AA p.f90fs*9 p.r248q CDS c.270delt c.743g>a breast Zygosity Homozygous Homozygous MDA-MB-468 AA p.? p.r273h CDS c.253+1g>t c.818g>a breast Zygosity Homozygous Homozygous PC3 AA p.r55fs*1 p.k139fs*31 CDS c.165_1212del1048 c.414delc prostate Zygosity Homozygous Homozygous RPMI-7951 AA p.? p.s166* p.v600e p.l16r CDS c.1_79del79 c.497c>a c.1799t>a c.47t>g melanoma Zygosity Homozygous Homozygous Heterozygous Homozygous SW1783 AA p.r233* p.r273h CDS c.697c>t c.818g>a glioma Zygosity Homozygous Heterozygous UM-UC3 AA p.m1_*404del p.f113c p.g12c p.m1_*157del CDS c.1_1212del1212 c.338t>g c.34g>t c.1_471del471 bladder Zygosity Homozygous Homozygous Homozygous Homozygous BT-474 AA p.k111n p.e285k CDS c.333g>c c.853g>a breast Zygosity Heterozygous Homozygous HEC1A AA p.g1049r p.r248q p.g12d CDS c.3145g>c c.743g>a c.35g>a endometrium Zygosity Heterozygous Homozygous Homozygous HCT116 AA p.h1047r p.g13d p.s252* p.r24fs*20 CDS c.3140a>g c.38g>a c.755c>a c.68_69insg colorectal Zygosity Heterozygous Heterozygous Homozygous Homozygous MCF7 AA p.e545k p.m1_*157del CDS c.1633g>a c.1_471del471 breast Zygosity Heterozygous Homozygous SUM-159 AA p.h1047l CDS c.3140a>t breast Zygosity Heterozygous MDA-MB-231 AA p.r280k p.g464v p.g13d p.m1_*157del CDS c.839g>a c.1391g>t c.38g>a c.1_471del471 breast Zygosity Homozygous Heterozygous Heterozygous Homozygous DU145 AA p.p223l, p.v274f p.? p.d84y CDS c.668c>t, c.820g>t c.117-2a>t c.250g>t prostate Zygosity Heterozygous Homozygous Homozygous HeLa AA CDS cervix Zygosity

Sample Name COSMIC Sample ID Amino Acid Nucleotide Primary Tissue Histology HCC70 907048 p.f90fs*9 c.270delt breast carcinoma 5839 cell-line S73189 850765 p.k60fs*9 c.180_181ins? breast carcinoma 9090379 5956 NS MDA- MB-468 850823 p.l70fs*7 c.208_251del44 breast carcinoma 9072974 4940 NS 895812 895812 p.a39fs*5 c.116_117insg central_nervous_system glioma 15517868 6230 S69959 849640 p.a79fs*20 c.237delc central_nervous_system glioma 10560660 5837 primary SKMG-3 847808 p.c71fs*6 c.212_255del44 central_nervous_system glioma 11515790 4919 NS S72911 850499 p.d24fs*20 c.70_71insa central_nervous_system glioma 9490783 5013 primary S75437 851274 p.d24fs*20 c.70_71insa central_nervous_system glioma 9393744 5013 primary S64444 845068 p.e43fs*11 c.127delg central_nervous_system glioma 9499454 5807 NS S70838 850036 p.f56fs*2 c.165_492del328 central_nervous_system glioma 10980610 5872 primary 895809 895809 p.k60* c.178a>t central_nervous_system glioma 15517868 6229 S75626 851448 p.k66fs*38 c.197_198ins14 central_nervous_system glioma 9619835 5022 primary S69259 849020 p.k6fs*4 c.17_18delaa central_nervous_system glioma 11438483 4929 primary S62854 844241 p.l70fs*7 c.210_253del44 central_nervous_system glioma 9426052 4887 primary S62857 844244 p.l70fs*7 c.210_253del44 central_nervous_system glioma 9426052 4887 primary SF539 905984 p.m1_*404del c.1_1212del1212 central_nervous_system glioma 14078 cell-line H4 907042 p.m1_*404del c.1_1212del1212 central_nervous_system glioma 14078 cell-line D- 245MG 946366 p.m1_*404del c.1_1212del1212 central_nervous_system glioma 14078 cell-line D- 336MG 946369 p.m1_*404del c.1_1212del1212 central_nervous_system glioma 14078 cell-line S69935 849616 p.n12fs*6 c.35_53del19 central_nervous_system glioma 10560660 4930 primary S68135 848253 p.r14fs*26 c.42_52del11 central_nervous_system glioma 12112531 4921 primary S73281 850849 p.r15fs*9 c.45_46insga central_nervous_system glioma 9072974 5014 primary A172 687563 p.r55fs*1 c.165_1212del1048 central_nervous_system glioma 26622 cell-line SW1088 909745 p.r55fs*1 c.165_1212del1048 central_nervous_system glioma 26622 cell-line U-87- MG 850822 p.v54fs*29 c.160_208del49 central_nervous_system glioma 9072974 4939 NS S64442 845066 p.y16fs*1 c.47_48insa central_nervous_system glioma 9499454 5879 NS S67801 847927 p.y46* c.138c>g central_nervous_system glioma 11939587 5307 primary Pubmed ID Mutation ID Sample Source

S65197 845784 p.y65* c.195c>a central_nervous_system glioma 10096247 5296 primary S69951 849632 p.y65* c.195c>g central_nervous_system glioma 10560660 5317 primary S69958 849639 p.y76fs*1 c.226_227delta central_nervous_system glioma 10560660 4963 primary S62656 844061 p.y76fs*1 c.227_228delat central_nervous_system glioma 9331071 4956 primary S68139 848257 p.y76fs*1 c.227_228delat central_nervous_system glioma 12112531 4956 primary S70960 850147 p.y88fs*3 c.263_264delat central_nervous_system glioma 9681833 4934 primary LS971 945577 p.y88fs*3 c.263_264delat central_nervous_system glioma 10949938 4934 NS S67198 847435 p.a3fs*14 c.9_30del22 endometrium carcinoma 11585417 4923 primary S65694 846198 p.c71fs*6 c.213_256del44 endometrium carcinoma 10657903 4905 primary S62930 844310 p.d24fs*19 c.71_72insc endometrium carcinoma 9887237 5880 primary S67536 847693 p.d24fs*19 c.71_72insc endometrium carcinoma 11006040 5880 primary S63111 844441 p.e18fs*5 c.54_57delggat endometrium carcinoma 9865913 4891 primary 1048066 1048066 p.e73fs*25 c.219_222delaaga endometrium carcinoma 17219201 23629 S75526 851354 p.e73fs*4 c.219_220delaa endometrium carcinoma 9307275 4948 primary 1009661 1009661 p.e7fs*3 c.19_20delga endometrium carcinoma 16021566 21341 - NOS 1048053 1048053 p.f56c c.167t>g endometrium carcinoma 17219201 23624 S75519 851347 p.f90fs*9 c.270delt endometrium carcinoma 9307275 5839 primary S68847 848789 p.h64fs*9 c.190_191delca endometrium carcinoma 11555573 4928 primary S63113 844443 p.i32fs*22 c.96delt endometrium carcinoma 9865913 5800 primary S66112 846569 p.i32fs*22 c.96delt endometrium carcinoma 9326929 5800 primary S63114 844444 p.i33fs*20 c.97_100delattg endometrium carcinoma 9865913 4893 primary S63078 844426 p.i33fs*21 c.97dela endometrium carcinoma 9829719 5798 primary S63079 844427 p.i33fs*21 c.97dela endometrium carcinoma 9829719 5798 metastasis E15857 738849 p.k6fs*4 c.17_18delaa endometrium carcinoma 12015762 4929 S73358 850914 p.k6fs*4 c.17_18delaa endometrium carcinoma 11849740 4929 primary 1048051 1048051 p.k6fs*4 c.17_18delaa endometrium carcinoma 17219201 4929 S67196 847433 p.l25fs*28 c.75_78delgacc endometrium carcinoma 11585417 4922 primary 1009628 1009628 p.l57fs*42 c.166_166delt endometrium carcinoma 16021566 21340 - NOS E15818 738809 p.l57fs*6 c.170_171inst endometrium carcinoma 12015762 5000

S68845 848787 p.l57fs*6 c.170_171inst endometrium carcinoma 11555573 5000 primary 1121036 1121036 p.l57fs*6 c.170_171inst endometrium carcinoma 18500270 5000 fixed - NOS S73359 850915 p.n48fs*6 c.143dela endometrium carcinoma 11849740 5834 primary S73362 850918 p.n63fs*10 c.187_188delaa endometrium carcinoma 11849740 4942 primary 1048052 1048052 p.n63fs*10 c.187_188delaa endometrium carcinoma 17219201 4942 1048067 1048067 p.n63fs*10 c.187_188delaa endometrium carcinoma 17219201 4942 S73361 850917 p.n63fs*11 c.188_189insa endometrium carcinoma 11849740 5017 primary S87002 852673 p.n63fs*36 c.187dela endometrium carcinoma 9354433 5836 primary S65506 846031 p.n63fs*36 c.188dela endometrium carcinoma 10746673 5811 primary S75527 851355 p.n63fs*36 c.188dela endometrium carcinoma 9307275 5811 primary S65507 846032 p.n63fs*36 c.188dela endometrium hyperplasia 10746673 5811 primary S65683 846187 p.n69fs*30 c.206dela endometrium carcinoma 10657903 5818 primary S63112 844442 p.p30fs*24 c.89delc endometrium carcinoma 9865913 5799 primary S86998 852669 p.r14fs*10 c.40dela endometrium carcinoma 9354433 5831 primary S65503 846028 p.r14fs*29 c.41_42insg endometrium carcinoma 10746673 5877 primary S67062 847299 p.r15fs*28 c.45_46inst endometrium carcinoma 11190805 5020 primary S68836 848778 p.r55fs*2 c.165_181del17 endometrium carcinoma 11555573 4926 primary S67546 847703 p.t26fs*28 c.78delc endometrium carcinoma 11006040 5833 primary S73360 850916 p.v45fs*10 c.134_135insca endometrium carcinoma 11849740 5016 primary S67493 847650 p.y16fs*21 c.46_65del20 endometrium carcinoma 11006040 4914 primary S82126 852119 p.y16fs*28 c.46_47inst endometrium carcinoma 15069681 5878 primary S77674 851776 p.y27fs*1 c.80_90del11 endometrium carcinoma 11156411 4964 primary S86999 852670 p.y27fs*16 c.80_81delat endometrium carcinoma 9354433 4952 primary 1048065 1048065 p.y29fs*25 c.83_83delt endometrium carcinoma 17219201 23628 S63056 844404 p.y68fs*5 c.202_203delta endometrium carcinoma 9829719 4889 primary S62932 844312 p.y76fs*1 c.227_228delat endometrium carcinoma 9887237 4956 primary S63078 844426 p.y76fs*1 c.227_228delat endometrium carcinoma 9829719 4956 primary OPM-2 909249 p.m1_*404del c.1_1212del1212 haematopoietic_and_lymphoid_tissue haematopoietic_neoplasm 14078 cell-line S70738 849939 p.m1fs*7 c.1_8delatgacagc haematopoietic_and_lymphoid_tissue lymphoid_neoplasm 9697884 5982 primary KE-37 907277 p.y27fs*1 c.80_1212del1133 haematopoietic_and_lymphoid_tissue haematopoietic_neoplasm 26623 cell-line 1141 1141074 p.e7fs*3 c.21_22delga large_intestine carcinoma 18781614 27631

980 1141072 p.f90fs*9 c.270delt large_intestine carcinoma 18781614 5839 1141 1141074 p.l57fs*42 c.170delt large_intestine carcinoma 18781614 5835 1190 1141075 p.l57fs*6 c.170_171inst large_intestine carcinoma 18781614 5000 S66226 846674 p.c83fs*9 c.247_248inst liver carcinoma 10363579 5883 primary H774 844562 p.? c.1-?_492+?del lung carcinoma 9696041 5779 NS NCI- H774 844809 p.? c.1-?_492+?del lung carcinoma 9598803 5779 NS SBC-1 713885 p.m1_*404del c.1_1212del1212 lung carcinoma 14078 cell-line NCI- H774 924243 p.m1_*404del c.1_1212del1212 lung carcinoma 14078 cell-line N230 844810 p.m1_?del fs c.1-?_79+?del lung carcinoma 9598803 5783 NS N231 844811 p.m1_?del fs c.1-?_79+?del lung carcinoma 9598803 5783 NS N417 844812 p.m1_?del fs c.1-?_79+?del lung carcinoma 9598803 5783 NS NCI- H1436 908469 p.r55fs*1 c.165_1212del1048 lung carcinoma 26626 cell-line LU-134- A 753588 p.y27fs*1 c.80_1212del1133 lung carcinoma 26623 cell-line S74259 851158 p.l57fs*42 c.170delt meninges meningioma 9549723 5835 primary 1131708 1131708 p.c71fs*28 c.213_213delt ovary carcinoma 17418409 27933 fresh - NOS - 979747 979747 p.f81fs*18 c.241_242tt>g ovary carcinoma 15668893 21619 NOS RTSG 909705 p.k6fs*4 c.17_18delaa ovary carcinoma 4929 cell-line S63057 844405 p.y68fs*5 c.202_203delta ovary carcinoma 9829719 4889 primary S63183 844508 p.g36fs*18 c.107delg prostate carcinoma 9823969 5802 primary S63185 844510 p.g44fs*11 c.131_139gcgtataca>acagaaagaca prostate carcinoma 9823969 5769 primary LNCaP- Clone- FGC 907788 p.k6fs*4 c.17_18delaa prostate carcinoma 4929 cell-line LNCaP 850745 p.k6fs*4 c.16_17delaa prostate carcinoma 9090379 4937 NS LNCaP 850821 p.k6fs*4 c.16_17delaa prostate carcinoma 9072974 4937 NS LNCaP 850413 p.k6fs*4 c.17_18delaa prostate carcinoma 9661880 4929 NS S72826 850417 p.q87* c.259c>t prostate carcinoma 9661880 5163 NS PC-3 905934 p.r55fs*1 c.165_1212del1048 prostate carcinoma 26622 cell-line

S72835 850426 p.y76fs*1 c.226_227delta prostate carcinoma 9661880 4963 NS FM95 851403 p.? c.1-?_492+?del skin malignant_melanoma 9288767 5779 NS S77773 851874 p.k6fs*4 c.17_18delaa skin malignant_melanoma 10651986 4929 metastasis G-mel 706101 p.m1_?del fs c.1-?_79+?del skin malignant_melanoma 10766161 5783 NS SW872 909756 p.m1_*404del c.1_1212del1212 soft_tissue liposarcoma 14078 cell-line UM-UC- 3 724838 p.m1_*404del c.1_1212del1212 urinary_tract carcinoma 14078 cell-line

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