Fig. 2

ZMP and ZTP accumulation in LND fibroblasts maintained with different culture media. A, B Fibroblasts were incubated for 5 days in RPMI medium with 2200 nM FA, 25 nM FA, 2200 nM FA + 350 μM UA or in HPLM medium. Cell extracts were obtained with 0.4 N PCA and ZMP and ZTP levels determined by HPLC. Results are expressed as nmol ZMP (A) or ZTP (B) per mg protein. C, D Fibroblasts were incubated for 7 days in Plasmax-PV medium containing 2.5% or 15% FBS and with 2200 nM or 25 nM FA. Cell extracts were obtained with 0.4 N PCA and ZMP and ZTP levels determined by HPLC. Results are expressed as nmol ZMP (C) or ZTP (D) per mg protein. E Doubling time was measured for fibroblasts cultivated for 7 days in each culture condition. Results are expressed as days. F Fibroblasts were incubated for 7 days in Plasmax-PV medium with 2200 nM or 25 nM FA and confluent fibroblasts were incubated only with Plasmax-PV containing 25 nM FA. Results are expressed as nmol ZMP per mg protein. G Doubling time of fibroblasts cultivated for 7 days in each culture condition. Results are expressed as days. Each graph represents the mean ± SEM of at least 3 control individuals and 3 patients with LND. *P < 0.05; P** < 0.01; P*** < 0.005 ****P < 0.001. Two-way ANOVA, Fisher’s multiple comparison test