Experiment Overview
| Repository ID: | FR-FCM-Z3ES | Experiment name: | G2/M transition following ATRi in Ccnc/Cdk8 WT/KO mESCs | MIFlowCyt score: | 96.42% |
| Primary researcher: | Rebecca Lloyd | PI/manager: | Steve Jackson | Uploaded by: | Rebecca Lloyd |
| Experiment dates: | 2018-09-07 - 2020-09-16 | Dataset uploaded: | Jan 2021 | Last updated: | Jan 2021 |
| Keywords: | [CCNC] [CDK8] [ATRi] [G2/M transition] | Manuscripts: | |||
| Organizations: |
University of Cambridge, Cambridge, (UK)
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| Purpose: | To assess whether loss of Ccnc or Cdk8 impact on the rate of transition through G2/M into the next cycle, in the presence and absence of ATR inhibition (ATRi). A reduction in this rate may explain why Ccnc/Cdk8 KO cells are resitant to ATRi. | ||||
| Conclusion: | As expected, ATRi treatment increased the rate of progression through mitosis, causing an increase in the proportion of cells in G1/S. The presence or absence of Ccnc or Cdk8 had no impact +/- ATRi. | ||||
| Comments: | Experimental protocol for EdU pulse-chase: 1. pulse cells with 10 uM EdU for 30 min. 2. wash cells twice with warm media 3. incubate (chase) for 6 h in DMSO or 900nM AZD6738 4. collect samples and stain for EdU. | ||||
| Funding: | Not disclosed | ||||
| Quality control: | A gating schematic including t=0 (30 min EdU and immediate collection) is also uploaded | ||||
Experiment variables
| Timepoints | |
|---|---|
| · 0h | 0h.fcs |
| · 6h | 021018 RL N2_WT 900nM 6h.fcs · 021018 RL N2_WT D1 900nM 6h.fcs · 021018 RL N2_WT D1 DMSO 6h 430.fcs · 021018 RL N2_WT DMSO 6h 520.fcs · 021018 RL N2_WT G2 900nM 6h.fcs · 021018 RL N2_WT G2 DMSO 6h 430.fcs · 070918_RL_WT 900nM 6h.fcs · 070918_RL_WT D1 900nM 6h.fcs · 070918_RL_WT D1 DMSO 6h.fcs · 070918_RL_WT DMSO 6h.fcs · 070918_RL_WT G2 900nM 6h.fcs · 070918_RL_WT G2 DMSO 6h.fcs · 6h ATRi CCNC D1.fcs · 6h ATRi CCNC G2.fcs · 6h ATRi CDK8 B9.fcs · 6h ATRi CDK8 H4.fcs · 6h ATRi WT.fcs · 6h DMSO CCNC D1.fcs · 6h DMSO CCNC G2.fcs · 6h DMSO CDK8 B9.fcs · 6h DMSO CDK8 H4.fcs · 6h DMSO WT.fcs · B9 900nM AZD6738.fcs · B9 DMSO.fcs · H4 900nM AZD6738.fcs · H4 DMSO.fcs · N2 120820 B9 900nM AZD6738.fcs · N2 120820 B9 DMSO.fcs · N2 120820 H4 900nM AZD6738.fcs · N2 120820 H4 DMSO.fcs · N2 120820 WT 900nM AZD6738.fcs · N2 120820 WT DMSO.fcs · N3 130820 B9 900nM AZD6738.fcs · N3 130820 H4 900nM AZD6738.fcs · N3 130820 H4 DMSO.fcs · N3 130820 WT 900nM AZD6738.fcs · N3 130820WT DMSO.fcs · N3130820 B9 DMSO.fcs · WT 900nM AZD6738.fcs · WT DMSO.fcs · 6h 900 nM AZD6738.fcs · 6h DMSO.fcs |
| Conditions | |
|---|---|
| · DMSO | 021018 RL N2_WT D1 DMSO 6h 430.fcs · 021018 RL N2_WT DMSO 6h 520.fcs · 021018 RL N2_WT G2 DMSO 6h 430.fcs · 070918_RL_WT D1 DMSO 6h.fcs · 070918_RL_WT DMSO 6h.fcs · 070918_RL_WT G2 DMSO 6h.fcs · 6h DMSO CCNC D1.fcs · 6h DMSO CCNC G2.fcs · 6h DMSO CDK8 B9.fcs · 6h DMSO CDK8 H4.fcs · 6h DMSO WT.fcs · B9 DMSO.fcs · H4 DMSO.fcs · N2 120820 B9 DMSO.fcs · N2 120820 H4 DMSO.fcs · N2 120820 WT DMSO.fcs · N3 130820 H4 DMSO.fcs · N3 130820WT DMSO.fcs · N3130820 B9 DMSO.fcs · WT DMSO.fcs · 0h.fcs · 6h DMSO.fcs |
| · 900 nM AZD6738 | 021018 RL N2_WT 900nM 6h.fcs · 021018 RL N2_WT D1 900nM 6h.fcs · 021018 RL N2_WT G2 900nM 6h.fcs · 070918_RL_WT 900nM 6h.fcs · 070918_RL_WT D1 900nM 6h.fcs · 070918_RL_WT G2 900nM 6h.fcs · 6h ATRi CCNC D1.fcs · 6h ATRi CCNC G2.fcs · 6h ATRi CDK8 B9.fcs · 6h ATRi CDK8 H4.fcs · 6h ATRi WT.fcs · B9 900nM AZD6738.fcs · H4 900nM AZD6738.fcs · N2 120820 B9 900nM AZD6738.fcs · N2 120820 H4 900nM AZD6738.fcs · N2 120820 WT 900nM AZD6738.fcs · N3 130820 B9 900nM AZD6738.fcs · N3 130820 H4 900nM AZD6738.fcs · N3 130820 WT 900nM AZD6738.fcs · WT 900nM AZD6738.fcs · 6h 900 nM AZD6738.fcs |
| Individuals | |
|---|---|
| · WT | 021018 RL N2_WT 900nM 6h.fcs · 021018 RL N2_WT DMSO 6h 520.fcs · 070918_RL_WT 900nM 6h.fcs · 070918_RL_WT DMSO 6h.fcs · 6h ATRi WT.fcs · 6h DMSO WT.fcs · N2 120820 WT 900nM AZD6738.fcs · N2 120820 WT DMSO.fcs · N3 130820 WT 900nM AZD6738.fcs · N3 130820WT DMSO.fcs · WT 900nM AZD6738.fcs · WT DMSO.fcs · 0h.fcs · 6h 900 nM AZD6738.fcs · 6h DMSO.fcs |
| · Ccnc KO (D1) | 021018 RL N2_WT D1 900nM 6h.fcs · 021018 RL N2_WT D1 DMSO 6h 430.fcs · 070918_RL_WT D1 900nM 6h.fcs · 070918_RL_WT D1 DMSO 6h.fcs · 6h ATRi CCNC D1.fcs · 6h DMSO CCNC D1.fcs |
| · Ccnc KO (G2) | 021018 RL N2_WT G2 900nM 6h.fcs · 021018 RL N2_WT G2 DMSO 6h 430.fcs · 070918_RL_WT G2 900nM 6h.fcs · 070918_RL_WT G2 DMSO 6h.fcs · 6h ATRi CCNC G2.fcs · 6h DMSO CCNC G2.fcs |
| · Cdk8 KO (B9) | 6h ATRi CDK8 B9.fcs · 6h DMSO CDK8 B9.fcs · B9 900nM AZD6738.fcs · B9 DMSO.fcs · N2 120820 B9 900nM AZD6738.fcs · N2 120820 B9 DMSO.fcs · N3 130820 B9 900nM AZD6738.fcs · N3 130820 H4 900nM AZD6738.fcs |
| · Cdk8 KO (H4) | 6h ATRi CDK8 H4.fcs · 6h DMSO CDK8 H4.fcs · H4 900nM AZD6738.fcs · H4 DMSO.fcs · N2 120820 H4 900nM AZD6738.fcs · N2 120820 H4 DMSO.fcs · N3 130820 H4 DMSO.fcs · N3130820 B9 DMSO.fcs |
| Sample Type | |
|---|---|
| · Biological replicate 1 | 070918_RL_WT 900nM 6h.fcs · 070918_RL_WT D1 900nM 6h.fcs · 070918_RL_WT D1 DMSO 6h.fcs · 070918_RL_WT DMSO 6h.fcs · 070918_RL_WT G2 900nM 6h.fcs · 070918_RL_WT G2 DMSO 6h.fcs |
| · Biological replicate 2 | 021018 RL N2_WT 900nM 6h.fcs · 021018 RL N2_WT D1 900nM 6h.fcs · 021018 RL N2_WT D1 DMSO 6h 430.fcs · 021018 RL N2_WT DMSO 6h 520.fcs · 021018 RL N2_WT G2 900nM 6h.fcs · 021018 RL N2_WT G2 DMSO 6h 430.fcs |
| · Biological replicate 3 | 6h ATRi CCNC D1.fcs · 6h ATRi CCNC G2.fcs · 6h ATRi CDK8 B9.fcs · 6h ATRi CDK8 H4.fcs · 6h ATRi WT.fcs · 6h DMSO CCNC D1.fcs · 6h DMSO CCNC G2.fcs · 6h DMSO CDK8 B9.fcs · 6h DMSO CDK8 H4.fcs · 6h DMSO WT.fcs |
| · Biological replicate 4 | N2 120820 B9 900nM AZD6738.fcs · N2 120820 B9 DMSO.fcs · N2 120820 H4 900nM AZD6738.fcs · N2 120820 H4 DMSO.fcs · N2 120820 WT 900nM AZD6738.fcs · N2 120820 WT DMSO.fcs |
| · Biological replicate 5 | N3 130820 B9 900nM AZD6738.fcs · N3 130820 H4 900nM AZD6738.fcs · N3 130820 H4 DMSO.fcs · N3 130820 WT 900nM AZD6738.fcs · N3 130820WT DMSO.fcs · N3130820 B9 DMSO.fcs |
| · Biological replicate 6 | B9 900nM AZD6738.fcs · B9 DMSO.fcs · H4 900nM AZD6738.fcs · H4 DMSO.fcs · WT 900nM AZD6738.fcs · WT DMSO.fcs |
| · Schematic | 0h.fcs · 6h 900 nM AZD6738.fcs · 6h DMSO.fcs |
