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. Author manuscript; available in PMC: 2015 Jun 1.
Published in final edited form as: Int J Obes (Lond). 2014 May 2;38(12):1578–1579. doi: 10.1038/ijo.2014.71

Figure 1.

Figure 1

Knockdown of the GR inhibits 3T3-L1 adipogenesis. Stable GR knockdown of 3T3-L1 preadipocyte lines were created with lentivirus delivery of short hairpin RNA (shRNA) targeted to the murine GR (GR shRNA) or a control, non-targeting shRNA (ctrl). After confirming GR knockdown, 3T3-L1 preadipocytes were differentiated following the standard protocol, induction in differentiation cocktail for 2 days (167 nM insulin, 1 μM dexamethasone, 0.5 mM IBMX in 10% fetal bovine serum (FBS) supplemented Dulbecco's modified Eagle's medium (DMEM)) and maintenance in DMEM+10% FBS. Cells were harvested and expression levels of GR and FABP4 (a maker of adipogenesis) were measured in cell lysates prepared before (d0) and after differentiation (d7). Data are from two independent GR shRNA sequences that showed more than 90% reduction in GR at the mRNA (a) and protein levels (b), and repeated three times with similar results.