ATR inhibition alone synergizes best with loss of ATR pathway genes, DNA replication genes, ERCC1, and ribonucleotide reductase. control (D) and REV3 knockdown cells (E and F). (G) Isobologram analysis of synergy. (H) Cells were treated with 1M ATRi, 0.1M cisplatin, or both (A + C); cells were released into press without medicines after 24 hours and allowed to form colonies. Error bars in all panels are standard deviation (n = 3).(TIF) pone.0125482.s002.tif (9.0M) GUID:?28A391F2-E17B-4F44-BEBE-AB4038E659EC S2 Fig: Related to Fig 7. Loss of REV3 is definitely synthetic lethal with ATRi and cisplatin. (A-F) A549 NSCLC cells were transfected with non-targeting siRNA (siNT) or two siRNAs focusing on REV3 (number 2 2 and 4 refer to specific sequences explained in the materials and methods). Cells were then treated with ATRi, cisplatin, and ATRi and cisplatin. Cell viability was identified with alamar blue and reported like a percent of the untreated control cells. (A) Level of sensitivity of REV3 knockdown cells to cisplatin. (B and C) Level of sensitivity of REV3 knockdown cells to ATRi and ATRi with 0.1M cisplatin. Bliss independence synergy between ATRi and cisplatin in control (D) and REV3 knockdown cells (E and F). Error bars in all panels are standard deviation (n = 3).(TIF) pone.0125482.s003.tif (5.8M) GUID:?D29038F0-4D8B-4705-A0ED-DBC4F5E18164 S3 Fig: Related to Fig 7. Loss of REV3 is definitely synthetic lethal with ATRi and cisplatin. (A-F) HCC1806 TNBC cells were transfected with non-targeting siRNA (siNT) or two siRNAs focusing on REV3 (number 2 2 and 4 refer to specific sequences explained in the materials and methods). Cells were then treated with ATRi, cisplatin, and ATRi and cisplatin. Cell viability was identified with alamar blue and reported like a percent of the untreated control cells. (A) Level of sensitivity of REV3 knockdown cells to cisplatin. (B and C) Level of sensitivity of REV3 knockdown cells to ATRi and ATRi with 0.1M cisplatin. Bliss independence synergy between ATRi and cisplatin in control (D) and REV3 knockdown cells (E and F). Error bars in all panels are standard deviation (n = 3).(TIF) pone.0125482.s004.tif (5.8M) GUID:?ABAE26A2-E523-4FC4-AEE1-3EF264769237 S4 Fig: Related to Fig 7. Loss of REV3 is definitely synthetic lethal with ATRi and cisplatin. (A-F) BT549 TNBC cells were transfected with non-targeting siRNA (siNT) or two siRNAs focusing on REV3 (number 2 2 and 4 refer to specific sequences explained in the materials and methods). Cells were then treated with ATRi, cisplatin, and ATRi and cisplatin. Cell viability was identified with alamar blue and reported like a percent of the untreated control cells. (A) Level of sensitivity of REV3 knockdown cells to cisplatin. (B and C) Level of sensitivity of REV3 knockdown cells to ATRi and ATRi with 0.5M cisplatin. Bliss independence synergy between ATRi and cisplatin in control (D) and REV3 knockdown cells (E and F). Error bars in all panels are standard deviation (n = 3).(TIF) pone.0125482.s005.tif (5.9M) GUID:?34C81AAB-C190-4EFA-BCE5-68EE54933EA5 S5 Fig: Related to Fig 8: Isobologram analysis of synergy in H157 using the dose response curve data in Fig 8. (TIF) pone.0125482.s006.tif (884K) GUID:?73C961AD-4B17-4463-8DC3-275BE57C48CF S6 Fig: Related to Fig 8: Loss of 53BP1 is definitely synthetic DDR1-IN-1 dihydrochloride lethal with ATRi and cisplatin. (A-F) A549 NSCLC cells were transfected with non focusing on siRNA (siNT) or two siRNAs focusing on 53BP1 (number 2 2 and 3 refer to specific sequences explained in the materials and methods). Cells were then treated with ATRi, cisplatin, and ATRi and cisplatin. Cell viability was identified with alamar blue and reported like a percent of the untreated control cells. (A) Level of sensitivity of 53BP1 knockdown cells to cisplatin. (B and C) Mapkap1 Level of sensitivity of 53BP1 knockdown cells to ATRi and ATRi with 0.5M cisplatin. Bliss independence synergy between ATRi and cisplatin in control (D) and 53BP1 knockdown cells (E and F). Error bars in all panels are standard deviation (n = 3).(TIF) pone.0125482.s007.tif (5.7M) GUID:?7C227B1D-7FFD-4700-8461-02D34458243E S7 Fig: Related to Fig 8: Loss of 53BP1 is definitely synthetic lethal with ATRi and cisplatin. (A-F) HCC1806 TNBC cells were transfected with non focusing on siRNA (siNT) or two siRNAs focusing on 53BP1 (number 2 2 and 3 refer to specific sequences explained in the materials and methods). Cells were then treated with ATRi, cisplatin, and ATRi and cisplatin. Cell viability was identified with alamar blue and reported like a percent of the DDR1-IN-1 dihydrochloride untreated control cells. (A) Level of sensitivity.The ATRi single-agent screen was previously published and included for comparison to the other drug treatments [18].(XLS) pone.0125482.s001.xls (387K) GUID:?A5C31675-CDCD-4458-A8F9-FCBD8D472801 S1 Fig: Related to Fig 7. and REV3 knockdown cells (E and F). (G) Isobologram analysis of synergy. (H) Cells were treated with 1M ATRi, 0.1M cisplatin, or both (A + C); cells were released into press without medicines after 24 hours and allowed to form colonies. Error bars in all panels are standard deviation (n = 3).(TIF) pone.0125482.s002.tif (9.0M) GUID:?28A391F2-E17B-4F44-BEBE-AB4038E659EC S2 Fig: Related to Fig 7. Loss of REV3 is definitely synthetic lethal with ATRi and cisplatin. (A-F) A549 NSCLC cells were transfected with non-targeting siRNA (siNT) or two siRNAs focusing on REV3 (number 2 2 and 4 refer to specific sequences explained in the materials and methods). Cells were then treated with ATRi, cisplatin, and ATRi and cisplatin. Cell viability was identified with alamar blue and reported like a percent of the untreated control cells. (A) Level of sensitivity of REV3 knockdown cells to cisplatin. (B and C) Level of sensitivity of REV3 knockdown cells to ATRi and ATRi with 0.1M cisplatin. Bliss independence synergy between ATRi and cisplatin in control (D) and REV3 knockdown cells (E and F). Error bars in all panels are standard deviation (n = 3).(TIF) pone.0125482.s003.tif (5.8M) GUID:?D29038F0-4D8B-4705-A0ED-DBC4F5E18164 S3 Fig: Related to Fig 7. Loss of REV3 is definitely synthetic lethal with ATRi and cisplatin. (A-F) HCC1806 TNBC cells were transfected with non-targeting siRNA (siNT) or two siRNAs focusing on REV3 (number 2 2 and 4 refer to specific sequences explained in the materials and methods). Cells were then treated with ATRi, cisplatin, and ATRi and cisplatin. Cell viability was identified with alamar blue and reported like a percent of the untreated control cells. (A) Level of sensitivity of REV3 knockdown cells to cisplatin. (B and C) Level of sensitivity of REV3 knockdown cells to ATRi and ATRi with 0.1M cisplatin. Bliss independence synergy between ATRi and cisplatin in control (D) and REV3 knockdown cells (E and F). Error bars in all panels are standard deviation (n = 3).(TIF) pone.0125482.s004.tif (5.8M) GUID:?ABAE26A2-E523-4FC4-AEE1-3EF264769237 S4 Fig: Related to Fig 7. Loss of REV3 is definitely synthetic lethal with ATRi and cisplatin. (A-F) BT549 TNBC cells were transfected with non-targeting siRNA (siNT) or two siRNAs focusing on REV3 (number 2 2 and 4 refer to specific sequences explained in the materials and methods). Cells were then treated with ATRi, cisplatin, and ATRi and cisplatin. Cell viability was identified with alamar blue and reported like a percent of the untreated control cells. (A) Level of sensitivity of REV3 knockdown cells to cisplatin. (B and C) Level of sensitivity of REV3 knockdown cells to ATRi and ATRi with 0.5M cisplatin. Bliss independence synergy between ATRi and cisplatin in control (D) and REV3 knockdown cells (E and F). Error bars in all panels are standard deviation (n = 3).(TIF) pone.0125482.s005.tif (5.9M) GUID:?34C81AAB-C190-4EFA-BCE5-68EE54933EA5 S5 Fig: Related to Fig 8: Isobologram analysis of synergy in H157 using the dose response curve data in Fig 8. (TIF) pone.0125482.s006.tif (884K) GUID:?73C961AD-4B17-4463-8DC3-275BE57C48CF S6 Fig: Related to Fig 8: Loss of 53BP1 is definitely synthetic lethal with ATRi and cisplatin. (A-F) A549 NSCLC cells were transfected with non focusing on siRNA (siNT) or two siRNAs focusing on 53BP1 (number 2 2 and 3 refer to specific sequences explained in the materials and methods). Cells were then treated with ATRi, cisplatin, and ATRi and cisplatin. Cell viability was identified with alamar blue and reported like a percent of the untreated control cells. (A) Level of sensitivity of 53BP1 knockdown cells to cisplatin. (B and C) Level of sensitivity of 53BP1 knockdown cells to ATRi DDR1-IN-1 dihydrochloride and ATRi with 0.5M cisplatin. Bliss independence synergy between ATRi and cisplatin in control (D) and 53BP1 knockdown cells (E and F). Error bars in all panels are standard deviation (n = 3).(TIF) pone.0125482.s007.tif (5.7M) GUID:?7C227B1D-7FFD-4700-8461-02D34458243E S7 Fig: Related to Fig 8: Loss of 53BP1 is definitely synthetic lethal with ATRi and cisplatin. (A-F) HCC1806 TNBC cells were transfected with non focusing on siRNA (siNT) or two siRNAs focusing on 53BP1 (number 2 2 and 3 refer to specific sequences explained in the materials and methods). Cells were then treated with ATRi, cisplatin, and ATRi and cisplatin. Cell viability was identified with alamar blue and reported like a percent of the untreated control cells. (A) Level of sensitivity of 53BP1 knockdown cells to cisplatin. (B and C) Level of sensitivity of 53BP1 knockdown.