Figure 5.

Differentiating 3T3-L1 cells responded heterogeneously to external perturbations. Differentiating 3T3-L1 cells were treated with pharmacological and metabolic perturbations for 24 h on D8. (a–c) The effects of either DMSO control or 10 µM troglitazone, as measured by population-averaged marker levels (a), 2D histograms of cell densities (b; triangles indicate centroids for S1–4 in the original subpopulation model; S_new indicates cells with previously unobserved phenotypes), and subpopulation-averaged PPARγ levels (c), are shown. (d) The effects of 12 perturbations were measured by absolute changes in population- and subpopulation-averaged marker levels and subpopulation distributions. Each perturbation had at least three replicates and 300 cells per replicate. (e) PPARγ levels in S4 versus PPARγ-binding abilities for the tested fatty acids and 15-d-PGJ2. Palmitic acid had a half-maximal inhibitory concentration value >30 µM and thus was excluded (ρ, Pearson’s correlation coefficient). Error bars indicate SEM (n = 3); *, P < 0.05; **, P < 0.01; ***, P < 0.001 by two-tailed t test.