Protocol tips
Upstream tips |
Protocol tips |
Downstream tips |
Always add your plasmid to the buffer before adding jetPrime |
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|
Upstream tips |
Always add your plasmid to the buffer before adding jetPrime |
Publication protocol
Luciferase assays
Primary chondrocytes were seeded at a density of 3 × 104 cells/well in 48 well plates 1 day prior to transfection. Cells were transfected with the pGL4.23-Mef2-luc (a gift from Eric Olson) or the pGL4.10-Ose2-luc reporter plasmid (a gift from G. Karsenty) in combination with pRL-TK (Promega) as a control and pCDNA3.1-DaCaMKII or pCDNA3.1-rKIIN expression vectors. Total amounts of transfected plasmids (equivalent to 52 fM) in each group were adjusted by adding empty vector (pCDNA3.1+). Transfection was performed using jetPRIME™ (Polyplus transfection) according to manufacturer's instructions. Luciferase activities were measured 48 h after transfection following the protocol by (Hampf and Gossen, 2006). Luciferase measurements were normalized to the corresponding renilla activities for transfection efficiency. Experiments were performed in triplicates and repeated at least three times.
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Papers
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Paper title
CaMKII Signaling Stimulates Mef2c Activity but Only Minimally Affects Murine Long Bone Development
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