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Ht31P toxicity
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Summary
- Stop stimulation
- Prepare washing buffer 1x ELISA + 10x PBS
- CoIP
Materials & Methods
Method
Preparations for ELISA & Viability staining
- Centrifuge sample incubated with beads, 5 min. 14.000 RPM
- Remove supernatant* (w. syringe)
- Add 1 mL PBS
- Centrifuge, 5 min. 14.000 RPM
- Remove supernatant*
- Add 75 μL Laemli buffer to beads
- Store samples @ -20 °C
* Be careful not to touch beads
Results
- PICTURES OF CELL DEATH (Anouk)
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Ht31P (μM)
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Ht31P (μM)
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Ht31P (μM)
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Ht31P (μM)
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Ht31P (μM)
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Ht31P (μM)
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D9 P23
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0
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10
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20
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30
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40
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50
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CTR
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7524 |
4117 |
2092 |
1929 |
1346 |
1712
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CTR
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6719 |
3030 |
1540 |
1323 |
1126 |
1458
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CSE
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3996 |
922 |
649 |
547 |
527 |
660
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CSE
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3509 |
1125 |
646 |
608 |
550 |
577
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Ht31P (μM)
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Ht31P (μM)
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Ht31P (μM)
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Ht31P (μM)
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Ht31P (μM)
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Ht31P (μM)
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D12 P27
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0
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10
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20
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30
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40
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50
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CTR
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7151 |
5242 |
3640 |
2953 |
2435 |
2331
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CTR
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6816 |
4353 |
2746 |
2339 |
1970 |
2074
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CSE
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5284 |
1784 |
1234 |
939 |
938 |
918
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CSE
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4858 |
1891 |
1250 |
1020 |
870 |
907
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Conclusion
- Ht31P is lethal for airway smooth muscle hTERT cells
- Strength of effect is donor dependent
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