Figure 3.

A20 protein is expressed constitutively in human MSCs. (a) A20 expression levels by real-time qPCR in MSCs, fibroblast and 293 T cells. Relative transcript expression levels were normalized to A20 transcript expression levels to human MSCs. (b) A20 protein analysis in three independent human bone marrow MSCs. Samples were compared with primary fibroblast from human skin and 293 T cell line. A20 protein expression level was measured by western blot using extracts where A20 was immunoprecipitated. (c) A20 expression during the differentiation process. A20 expression levels were measured by western blot in total cell extracts using the antibody for A20. Sam68 was used as a loading control (d) A20 expression is dependent of NF-κB activation. IKKβ (MSC-IKKi) and IκBα (MSC- IκBi) transcripts were ‘knocked down' by specific shRNA. Six-days post-infection MSCs were analyzed for A20 expression levels by real-time qPCR. Relative transcript expression levels were normalized to MSCs transduced with the empty vector (MSC-pLV)