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Using these plasmids as templates, PCR was performed to modify the N-terminus of the expressed protein to enhance protein expression as described previously [6].
Correct immunophenotype was confirmed by flow cytometry for B220 expression as described above.
Ig sequences were transfected into HEK293 cells (Expi293F™ Cells, Gibco, Cat. No. A14635) for antibody expression as described below.
The FinProg series (1,212 samples) was previously stained for pSrcY416 and MYO10 expression as described earlier 13,32
The lc_sigma_clip routine can create a GTI file directly, or you can use the limits it calculates to create the file yourself using the dmgti tool, or use them directly within a DM filter expression, as described below.
Control BST transfection was determined by staining for GFP expression, as described in [13].
After return to a resting phase, cells were analysed for cytokine expression, as described above.
We quantified egr-1 expression as described in Burmeister et al. 2008 [2008
All plasmids were sequence verified and tested for correct expression as described below.
Relative expression (V) was calculated by normalization to ß-actin expression as described in [26].
Cells were lysed and immunoblotted for PI3K expression as described before [89].
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