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. 2016 Sep 23;155(1):85–100. doi: 10.1093/toxsci/kfw186

FIG. 2.

FIG. 2

Optimization of a PPARG-dependent human adipogenesis assay. A donor pool of non-differentiated adipose-derived stem cells was tested for adipogenic capacity and expression of endogenous PPARG isoforms at the end of the 8 day assay protocol. A Western blot of non-differentiated (Neg Con) and differentiated (Rosi 1 µM) cells was probed for PPARG, FABP4, and GAPDH expression (A). A modified medium formulation illustrates concentration-responsive differentiation of rosiglitazone (B) and tributyltin (C) treated cells, multiplexed with acute (LDH release) and chronic (cell counts) cytotoxicity endpoints. Concentrations are mean ± SEM of 3 experimental replicates. Micrographs taken with a 20× objective demonstrate brightfield (BF), nuclear (Hoechst), neutral lipid (AdipoRed), and composite images of the 4 control samples used in the adipogenesis screen (D). Neg Con = non-differentiated hASC, DMSO= vehicle, Rosi= rosiglitazone, TBT= tributyltin.