Fig 11.

Adipogenesis is impaired in HuR-depleted wild-type MEF. (A) Wild-type MEFs were infected with sh-SCR or sh-HuR retroviruses. Selected cells were stimulated to differentiate them into adipocytes in the presence of insulin, 3-isobutyl-1-methylxanthine (IBMX), and dexamethasone for 10 days and stained with Oil Red O. Bars, 50 μm. (B) Wild-type MEFs with sh-SCR or sh-HuR were cultured in adipocyte differentiation medium for the indicated periods. The expression of C/EBPα, -β, -δ, and PPARγ were analyzed by immunoblotting. (C) ARF-null MEFs with sh-SCR or sh-HuR were stimulated to differentiate them for 10 days and stained with Oil Red O. Bars, 50 μm. (D and E) Wild-type (D) and ARF-null (E) MEFs expressing sh-SCR or sh-HuR were stimulated to differentiate them for 0, 1, and 2 days. Cells were pulse-labeled with EdU for 45 min and stained for EdU. EdU-positive and -negative cells in microscopic fields were counted. Data are representative of two independent experiments. Error bars represent SEM of results from five microscopic fields.