Fig. 8.

Human fibroblast populations derived from patients carrying biallelic LPIN1 inactivating mutations can be induced to adipogenic differentiation. Human dermal primary fibroblast populations derived from normal control (healthy) individuals or from lipin-1-defective patients (see Table 1) were cultured until confluence in GM. Cells were then shifted to adipogenic differentiation medium, DM (+), containing insulin, dexamethasone, isobutylmethylxanthine, and indometacin, or left in GM (−). After 4, 8, or 12 days in DM, cells were harvested, the total RNA extracted, and first-strand cDNA generated from 1 μg of total RNA to perform real-time PCR using specific primers for PPARG (A), FASN (B), and PGC1A (C). Expression data were normalized using actin β (ACTB) as a housekeeping gene. Relative expression values are expressed as mean values ± SD.