Additional file 10.
Dynamic change of neurofibromin level in lymphoblastoid cell lines in response to serum deprivation. We determined the effects of serum deprivation on neurofibromin level as a way to establish the physiologic relevance of lymphoblastoid cell lines (LCLs) in the study of NF1-haploinsufficiency. We measured levels of neurofibromin in two LCLs (one each from NF1-affected, and -unaffected individuals) that were serum-deprived (0.1% serum) for 16 hours, and then released for variable amounts of time in complete (10% serum; supports cell proliferation), or incomplete (1% serum; does not support cell proliferation) medium. Western blot analysis and quantitation of relative abundance of neurofibromin in an NF1-unaffected individual (A) and an NF1-affected individual (B). NF1 abundance data is shown to the right from respective western blot and is plotted as percentage relative to NF1 abundance in exponentially growing cells. "E" - exponentially growing LCLs; "S" - serum starved LCLs; 5', 30', 7 h - cells released into media containing either 1% or 10% FBS for 5 min, 30 min or 7 hours, respectively. Our experiment showed that the amount of neurofibromin increased approximately two-fold in serum-starved cells as compared to that in exponentially growing LCLs from both affected and unaffected individuals. When the cells were released into complete medium (10% FBS), the neurofibromin level quickly returned to pre-starvation levels in both NF1-affected and -unaffected LCLs. In contrast, the neurofibromin level continued to increase during prolonged incubation of the cells in incomplete medium (1% FBS). We conclude that in LCLs neurofibromin level is sensitive to environmental conditions.
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Pemov et al. BMC Genomics 2010 11:194 doi:10.1186/1471-2164-11-194