User:Karmella Haynes/Notebook/Polycomb project/2010/06/28

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06/28/10

 * &#x2713; Senescence assay: C12-FDG treatment & flow cytometry

Senescence assay > Samples (~3.5 mL medium/ well, plated on 6/24/20): > Note: added fresh dox to "dox" samples on 6/27 > C12-FDG: add 30 μg/mL; 37°C/ 1 hr.; harvest and strain cells for flow cytometry (1x106 cells/ 500 μL) > Flow cytometry: RFP and FITC channels, count 10,000 live cells per sample
 * 1) KAH126-1
 * 2) KAH126-1 +1 μg/mL dox
 * 3) KAH126-3
 * 4) KAH126-3 +1 μg/mL dox
 * 5) KAH132-8
 * 6) KAH132-8 +1 μg/mL dox
 * 7) KAH154-2
 * 8) KAH154-2 +1 μg/mL dox
 * 9) FTRx + 0 rotenone (7.0 μL DMSO)
 * 10) FTRx + 0.1 μg/mL rotenone (1.75 μL/ 5.25 μL DMSO)
 * 11) FTRx + 0.2 μg/mL rotenone (3.5 μL/ 3.5 μL DMSO)
 * 12) FTRx + 0.4 μg/mL rotenone (7.0 μL/ 0 μL DMSO)
 * 13) FTRx blank

> Results:
 * Background signal too high (104); titer C12-FDG and rotenone to lower concentrations for FTRx cells
 * Induction of RFP successful for all Pc-ATF cells. Off-state nearly equal to FTRx untransfected cells. 126-1, 126-3 show a sub-population of non-induced cells after dox treatment.


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