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Controls were performed with cells maintained in medium with 5% heat inactivated FCS for 6 h.
Single cell clones were maintained in medium with 10 µg/ml puromycin.
To avoid prestimulation by serum withdrawal, primary fibroblasts were maintained in medium with 0.5% FCS prior to stimulation with TNF.
After 72 hours, positive clones were selected in growth medium containing G418 (0.4 mg/ml) and resistant cells were maintained in medium with G418 (0.2 mg/ml).
Transcription of c-KIT and both SCF splice variants was active in normoxic controls and cells subjected to hypoxia followed by a 24 hour recovery while maintained in medium with or without recombinant rat (rr) SCF (100 ng/ml) (figure 6C).
After selection, cells were maintained in medium with 400 μg/ml G-418.
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Stable cell lines with Jak2 in pSG213 were maintained in medium, supplemented with puromycin (1 µg/ml), and stable cell lines with wtGHR with hygromycin 50 µg/ml).
Cells were washed with PBS and maintained in medium free with PTD-poMx1 for 48 h at 37 °C.
Cells were washed three times with PBS and maintained in medium free with PTD-poMx1 for 48 h at 37 °C.
They were maintained in medium supplemented with E2.
Cells were maintained in medium supplemented with 1 × penicillin and streptomycin (Invitrogen).
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