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Images of wounded cells were taken immediately and 24 h later after incubation in SABM media with supplements (Olympus IX70 microscope; Olympus, Center Valley, PA).
16C (ME16C) [ 61] were cultured in HuMEC media with supplements (Gibco).
FAC-sorted cell fractions were placed on HuMEC media with supplements, 5 % FBS and 5 % Matrigel™.
EGM-2 MV SingleQuots Kit media with supplements and growth factors (Lonza) were used for ECs.
A549 cells were grown in Hams F12 media with supplements as above.
SUM149PT was maintained in HuMEC media with supplements (Gibco) with 5 % FBS [ 62].
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Before the collection of media for secretion analysis, all media with supplement were withdrawn and the cells were washed 5 times with the differentiation media (without supplement).
An EBV transformed B Cell Line (BCL) was cultured in RPMI media (Sigma) with supplements.
Cells were serum starved overnight in DMEM media with no supplements.
After imaging selected cells for several minutes, we replaced the growth media with media supplemented with 2 nM Latrunculin B. This concentration has been used in the past to reversibly terminate growth [2], [39].
The inserts were assembled into 24-well plates with each well containing 700 μl of media with 10% supplemented with fetal calf serum, which was used as chemoattractant.
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