Experiment Overview
| Repository ID: | FR-FCM-Z5XQ | Experiment name: | Escape from G1 arrest during acute MEK inhibition drives the acquisition of drug resistance | MIFlowCyt score: | 56.00% |
| Primary researcher: | Prasanna Channathodiyil | PI/manager: | Jon Houseley | Uploaded by: | Prasanna Channathodiyil |
| Experiment dates: | 2018-12-13 - 2020-12-19 | Dataset uploaded: | Sep 2022 | Last updated: | Sep 2022 |
| Keywords: | [EdU incorporation] [[5-ethynyl-2’-deoxyuridine]] [Cyclin B1 staining] | Manuscripts: | |||
| Organizations: |
Babraham Institute, Babraham Institute, Cambridge, Cambridge (United Kingdom)
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| Purpose: | To assess whether Selumetinib-treated colorectal cancer cells escape G1 cell cycle arrest using incorporation assays for the thymidine analogue 5- ethynyl 2-deoxyuridine (EdU) and to characterise cells that undergo DNA replication during short-term and/or prolonged treatment with Selumetinib | ||||
| Conclusion: | Selumetinib causes G1 cell cycle arrest, but cells occasionally enter the cell cycle and initiate DNA replication despite robust G1 cell cycle arrest induced by the drug. Combined treatment with selumetinib and a dose of palbociclib sufficient to reinforce G1 arrest in selumetinib-sensitive cells, but not to impair proliferation of resistant cells, delays the emergence of resistant colonies | ||||
| Comments: | None | ||||
| Funding: | This study was funded by Wellcome Trust [110216] and the BBSRC [BI Epigenetics ISP: 399 BBS/E/B/000C0423] | ||||
| Quality control: | Appropriate control cell populations were included for each assay. For each EdU incorporation assay, control cells that were unstained for EdU and underwent the same incubation and washing steps were used | ||||
Experiment variables
| Sample Type | |
|---|---|
| · COLO205 cells | Specimen_001_Clone 1 - Tra 3nm 24h + Edu 4h_004.fcs · Specimen_001_Clone 1 Control- Log_002.fcs · Specimen_001_Clone 1 unstained_001.fcs · Specimen_001_Clone 2 Palbo-R1_011.fcs · Specimen_001_Clone 2 Palbo-R2_012.fcs · Specimen_001_Clone 2 Sel+Palbo-R1_023.fcs · Specimen_001_Clone 2 Sel+Palbo-R2_024.fcs · Specimen_001_Clone 2 Sel-R1_017.fcs · Specimen_001_Clone 2 Sel-R2_018.fcs · Specimen_001_Clone 2 untreated-R1_005.fcs · Specimen_001_Clone 2 untreated-R2_006.fcs · Specimen_001_Clone 3 Treated_009.fcs · Specimen_001_Clone 3 Untreated_008.fcs · Specimen_001_Clone 6 Log+Edu_002.fcs · Specimen_001_Clone 6 Sel+Edu_003.fcs · Specimen_001_Clone 6 Treated_015.fcs · Specimen_001_Clone 6 Untreated_014.fcs · Specimen_001_Control Treated_003.fcs · Specimen_001_Control Untreated_002.fcs · Specimen_001_Control2_004.fcs · Specimen_001_Parental Palbo-R1_009.fcs · Specimen_001_Parental Palbo-R2_010.fcs · Specimen_001_Parental Sel+Palbo-R1_021.fcs · Specimen_001_Parental Sel+Palbo-R2_022.fcs · Specimen_001_Parental Sel-R1_015.fcs · Specimen_001_Parental Sel-R2_016.fcs · Specimen_001_Parental untreated-R1_003.fcs · Specimen_001_Parental untreated-R2_004.fcs · Specimen_001_S1 R1_005.fcs · Specimen_001_S1 R2_006.fcs · Specimen_001_S1 R3_002.fcs · Specimen_001_S2 R1_007.fcs · Specimen_001_S2 R2_008.fcs · Specimen_001_S2 R3_003.fcs · Specimen_001_S3 R1_009.fcs · Specimen_001_S3 R2_010.fcs · Specimen_001_S3 R3_004.fcs · Specimen_001_S4 R1_011.fcs · Specimen_001_S4 R2_012.fcs · Specimen_001_S4 R3_005.fcs · Specimen_001_Sel no EdU R1_003.fcs · Specimen_001_Sel no EdU R2_004.fcs · Specimen_001_Sel1 R1_005.fcs · Specimen_001_Sel1 R2_006.fcs · Specimen_001_Sel2 R1_007.fcs · Specimen_001_Sel2 R2_008.fcs · Specimen_001_Sel3 R1_009.fcs · Specimen_001_Sel3 R2_010.fcs · Specimen_001_Sel4 R1_011.fcs · Specimen_001_Sel4 R2_012.fcs · Specimen_001_Sel5 R1_013.fcs · Specimen_001_Sel5 R2_014.fcs · Specimen_001_SelnoEdUR1_003.fcs · Specimen_001_SelnoEdUR2_004.fcs · Specimen_001_Tube_001_001.fcs · Specimen_001_Tube_002_002.fcs · Specimen_001_Tube_003_003.fcs · Specimen_001_Tube_004_004.fcs · Specimen_001_Tube_005_005.fcs · Specimen_001_Tube_006_006.fcs · Specimen_001_Tubes_001_001.fcs · Specimen_001_Tubes_002_002.fcs · Specimen_001_Tubes_003_003.fcs · Specimen_001_Tubes_004_004.fcs · Specimen_001_Tubes_005_005.fcs · Specimen_001_Tubes_006_006.fcs · Specimen_001_Tubes_007_007.fcs · Specimen_001_Unstained + DAPI_001.fcs · Specimen_001_Unstained_001.fcs · Specimen_001_control-2_005.fcs · Specimen_001_no EdU control_002.fcs · Specimen_001_treated-2_004.fcs · Specimen_001_treated_003.fcs · Specimen_001_unstained+DAPI_001.fcs · Specimen_001_unstained_.fcs · Specimen_001_Colo unstained_001.fcs |
| · HT29 cells | Round 2_Sel - R1_011.fcs · Round 2_Unstained + DAPI R1_007.fcs · Round 2_Untreated - R1_009.fcs |
| Conditions | |
|---|---|
| · EdU stained | Round 2_Sel - R1_011.fcs · Round 2_Untreated - R1_009.fcs · Specimen_001_Clone 1 - Tra 3nm 24h + Edu 4h_004.fcs · Specimen_001_Clone 1 Control- Log_002.fcs · Specimen_001_Clone 2 Palbo-R1_011.fcs · Specimen_001_Clone 2 Palbo-R2_012.fcs · Specimen_001_Clone 2 Sel+Palbo-R1_023.fcs · Specimen_001_Clone 2 Sel+Palbo-R2_024.fcs · Specimen_001_Clone 2 Sel-R1_017.fcs · Specimen_001_Clone 2 Sel-R2_018.fcs · Specimen_001_Clone 2 untreated-R1_005.fcs · Specimen_001_Clone 2 untreated-R2_006.fcs · Specimen_001_Clone 3 Treated_009.fcs · Specimen_001_Clone 3 Untreated_008.fcs · Specimen_001_Clone 6 Treated_015.fcs · Specimen_001_Clone 6 Untreated_014.fcs · Specimen_001_Control Treated_003.fcs · Specimen_001_Control Untreated_002.fcs · Specimen_001_Parental Palbo-R1_009.fcs · Specimen_001_Parental Palbo-R2_010.fcs · Specimen_001_Parental Sel+Palbo-R1_021.fcs · Specimen_001_Parental Sel+Palbo-R2_022.fcs · Specimen_001_Parental Sel-R1_015.fcs · Specimen_001_Parental Sel-R2_016.fcs · Specimen_001_Parental untreated-R1_003.fcs · Specimen_001_Parental untreated-R2_004.fcs · Specimen_001_S1 R1_005.fcs · Specimen_001_S1 R2_006.fcs · Specimen_001_S1 R3_002.fcs · Specimen_001_S2 R1_007.fcs · Specimen_001_S2 R2_008.fcs · Specimen_001_S2 R3_003.fcs · Specimen_001_S3 R1_009.fcs · Specimen_001_S3 R2_010.fcs · Specimen_001_S3 R3_004.fcs · Specimen_001_S4 R1_011.fcs · Specimen_001_S4 R2_012.fcs · Specimen_001_S4 R3_005.fcs · Specimen_001_Sel1 R1_005.fcs · Specimen_001_Sel1 R2_006.fcs · Specimen_001_Sel2 R1_007.fcs · Specimen_001_Sel2 R2_008.fcs · Specimen_001_Sel3 R1_009.fcs · Specimen_001_Sel3 R2_010.fcs · Specimen_001_Sel4 R1_011.fcs · Specimen_001_Sel4 R2_012.fcs · Specimen_001_Sel5 R1_013.fcs · Specimen_001_Sel5 R2_014.fcs · Specimen_001_Tube_002_002.fcs · Specimen_001_Tube_003_003.fcs · Specimen_001_Tube_004_004.fcs · Specimen_001_Tube_005_005.fcs · Specimen_001_Tube_006_006.fcs · Specimen_001_Tubes_002_002.fcs · Specimen_001_Tubes_003_003.fcs · Specimen_001_Tubes_004_004.fcs · Specimen_001_Tubes_005_005.fcs · Specimen_001_Tubes_006_006.fcs · Specimen_001_Tubes_007_007.fcs · Specimen_001_control-2_005.fcs · Specimen_001_treated-2_004.fcs |
| · Unstained | Round 2_Unstained + DAPI R1_007.fcs · Specimen_001_Clone 1 unstained_001.fcs · Specimen_001_Sel no EdU R1_003.fcs · Specimen_001_Sel no EdU R2_004.fcs · Specimen_001_SelnoEdUR1_003.fcs · Specimen_001_SelnoEdUR2_004.fcs · Specimen_001_Tube_001_001.fcs · Specimen_001_Tubes_001_001.fcs · Specimen_001_Unstained + DAPI_001.fcs · Specimen_001_Unstained_001.fcs · Specimen_001_no EdU control_002.fcs · Specimen_001_unstained+DAPI_001.fcs · Specimen_001_unstained_.fcs · Specimen_001_Colo unstained_001.fcs |
| · EdU and Cyclin B1 co-stained | Specimen_001_Clone 6 Log+Edu_002.fcs · Specimen_001_Clone 6 Sel+Edu_003.fcs |
| · Cyclin B1 staining | Specimen_001_Control2_004.fcs · Specimen_001_treated_003.fcs |
