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Fig. 2. ADAM12 mRNA expression is transiently upregulated before adipocyte differentiation. (A) Total RNA was extracted from mouse 3T3-L1 cell cultures at various times during differentiation. The levels of ADAM12, PPAR{gamma} and GAPDH mRNA expression were analyzed by northern blot. GAPDH and rRNA served as loading controls. (B) Quantitative analysis of ADAM12 and PPAR{gamma} mRNA expression, normalized to GAPDH mRNA levels. (C) Northern blot analysis of ADAM12 RNA extracted from human liposarcoma-derived LiSa-2 cells at various stages of differentiation. (D) ADAM12 protein expression was analyzed by western blot in extracts from growing C3H10T1/2, 3T3-L1 and LiSa-2 cells. CHOK1 cells transfected with control vector and full-length mouse ADAM12 cDNA were used as negative (NC) and positive (PC) controls, respectively. D0, day 0 confluent cells; D3, D4, D9 and D17, days 3, 4, 9 and 17 after the onset of differentiation, respectively; G, growing fibroblastic cells.