New Transcriptional Reporters to Quantify and Monitor PPARγ Activity

Figure 2

PPRE-H2B-eGFP and PPRE-pNL1.3[secNluc] transfections of VCT: new models to visualize and quantify the activity of PPARγ. (a) Merged staining of PPRE-H2B-eGFP (green), cell shape using F-actin (red), and nuclei using DAPI (blue) in 48 h treated VCT with 1 μM GW1929 (agonist of PPARγ) or 1 μM GW9662 (PPARγ antagonist) compared to control (vehicle). 3D nuclei segmentation (b) and quantification (c) of PPRE-H2B-eGFP fluorescence intensity using Icy software. (d) PPRE-H2B-eGFP protein levels were assessed using western blotting. (e-f) Spectrophotometry reading of luciferase expression for 50 μl (e) or 100 μl (f) of supernatant from 24- and 48-hour PPRE-pNL1.3 transfected and 1 μM GW1929 or 1 μM GW9662 treated VCT compared to nontransfected cells (control). Values are represented as mean ± SD; < 0.001, < 0.0001 versus vehicle control ().

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