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Previous data on VEGF regulation has been obtained using cell lines; we demonstrate our phenomenon in primary human monocytes, enhancing the relevance of this observation.
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Hence all data presented here were obtained using cell suspensions and non-invasive techniques.
The DNA content was analyzed with FACS vantage SE BD Biosciencess), and the data were obtained using Cell Quest Software (BD Biosciences).
Protein extracts were obtained using Cell Extraction Buffer buffer (BioSource International, CA, USA) plus protease inhibitor cocktail (Sigma).
Similar results were obtained using cell extracts expressing GFP-tagged domain II of TPC1.
Briefly, total cell lysates were obtained using cell lysis buffer (Sigma-Aldrich) on ice for 10 min.
Maximal cell migration (chemotaxis) was obtained using cell migration media containing ten (10) percent fetal calf serum (FCS).
As many previous results were obtained using cell lines cultured in vitro, they may not reflect dynamic changes of histone modifications occurring during differentiation in vivo.
Similar results were obtained using cell-based assays, and in B16F10 cells, compound 1m dose-dependently inhibited tyrosinase activity and melanogenesis.
Similar results were obtained using cells stably transfected with Myc hVps34 and V5 hVps15.
Essentially similar results were obtained using cells expressing domain II encoded by TPC1427 816 Myc.
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