Manual - New England Biolabs

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Luciferase Activity (RLU) 10 100,000 10,000 1,000 100 10 1 RSL1 (500 nM) DMSO 0 4 8 12 16 20 24 Time Post Induction (Hours) Figure 5: Induction time course. NIH 3T3 cells co-transfected with pNEBR-R1 and pNEBR- X1GLuc control plasmid were induced with RSL1. Gaussia luciferase activity was assayed between 1 and 24 hours post-induction.
Luciferase Activity (RLU) 25,000 20,000 15,000 10,000 5,000 RSL1 Addition RSL1 (500 nM) DMSO Control RSL1 Removal RSL1 Addition RSL1 Removal 0 0 12 24 36 48 Hours 60 72 84 96 Figure 6: Rapid ON:OFF switch response to two cycles of addition and removal of RSL1. NIH 3T3 cells were co-transfected with pNEBR-R1 and pNEBR-X1 expressing a destabilized firefly luciferase and induced with RSL1 followed by removal of RSL1 at 24 hours. A second cycle of addition/removal of RSL1 results in a new ON:OFF switch response. To remove RSL1 and turn off expression, cells were washed and RSL1-free media was added at the times indicated. The blue line represents induced expression. The red line represents uninduced expression. 11
Page 1: Catalog #E3000S Store at -20°C Rhe
Page 4 and 5: 2 Supplied Components: The RheoSwit
Page 6 and 7: 4 Introduction: The RheoSwitch ® M
Page 8 and 9: 6 amounts of RSL1 (Figure 4). Even
Page 10 and 11: 8 Luciferase Activity (RLU) 90,000
Page 14 and 15: 12 Gaussia Luciferase (GLuc) Contro
Page 16 and 17: 14 To induce expression, RSL1 ligan
Page 18 and 19: 16 4. Add RSL1 ligand to a final co
Page 20 and 21: 18 B. PshA I 3640 BfuA I - BspM I 3
Page 22 and 23: 20 References: 1. Karzenowski, D.,
Page 24: 22 Licensing Information: RheoSwitc

Manual - New England Biolabs

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