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For obtaining hypermethylated regions, we supplied late passage as signal and early passage as control signal.
PCNA staining confirmed lack of proliferation in late passage cells under static or flow conditions.
Late passage EC do not proliferate in response to turbulent flow as younger EC do.
We designed an in vitro screening platform by using late passage (passage 5) of WRN -/- MSCs at low confluence.
Late passage EC also do not phosphorylate Akt, and attract fewer SMC, than early passage EC, in response to flow.
Western blotting demonstrated increased Akt phosphorylation with flow in early, but not late, passage EC (n = 6, P = 0.01).
Cultures of late passage mOP cells grown in proliferation medium are highly pure early stage oligodendrocyte precursors where >90% assume a characteristic bipolar morphology.
Early passage cells demonstrated increased proliferation with flow compared to static conditions (36% increase, day 5, P = 0.03); late passage cells had no statistically significant increase (P = 0.42).
Late passage (passage 270, L) represents cells that underwent about 1000 doublings.
Similar(2)
HMECs from early and late passages displayed a similar and stable methylome, as did early- and late-passage vHMECs.
As expected, a marked reduction of WFDC1 mRNA level in the late passage-derived samples was observed in both systems.
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