Figure 6
From: Identification of the NF-κB inhibitor A20 as a key regulator for human adipogenesis
A20 overexpression stimulates the adipogenic capacity of human MSCs. (a) MSCs were transduced with the empty bicistronic expression vector pRRL (MSC-pRRL) or containing the A20 cDNA (MSC-A20). Transduction efficiency was analyzed by flow cytometry (GFP+ cells). A20 overexpression was determined by western blot in total cell extracts. Sam68 was used as a loading control. (b) Quantification by real-time qPCR of IL-6. MSCs overexpressing A20 transcript for 4 days were cultured for additional 24 h in the presence or absence of 15 ng/ml TNFα. Relative transcript IL-6 expression levels were normalized to MSCs transduced with the empty vector cultured in the absence of TNFα. (c) MSCs differentiation capacity is influenced by A20 overexpression. MSCs showing ∼50% infection efficiency were induced to differentiate into adipocytes in the presence or absence of TNFα. Percentages of mature adipocytes from GFP− (white bars) and GFP+ (gray bars) cells (with the empty vector or A20-cDNA expressing vector, respectively) after the induction period are represented. Results show the mean±S.D. of triplicates samples. ***P<0.001
