Fig. 2: NCT and NFT are HNF4α agonists. | Cell Death & Disease

Fig. 2: NCT and NFT are HNF4α agonists.

From: Liver fat storage is controlled by HNF4α through induction of lipophagy and is reversed by a potent HNF4α agonist

Fig. 2

A HNF4α siRNA blocked the effect of HNF4α agonists. Scrambled or HNF4α siRNAs were transfected into T6PNE cells 2 days before compound administration. Compounds at the indicated concentrations were treated for an additional 2 days. The cells were analyzed for the percentage of cells expressing the insulin promoter-GFP transgene (N = 6). B DARTS assay to detect effect of compounds on HNF4α protease sensitivity. For the DARTS assay, HepG2 cells were treated with DMSO (lane 1), BI6015 (lane 2), NCT (lane 3), or NFT (lane 4) at a concentration of 40 or 80 μM for 16 h. Total cell protein was extracted and each sample was split into two aliquots for proteolysis without (−) or with (+) subtilisin and analyzed by Western blotting for HNF4α as done previously7. After detection of HNF4α, the membrane was stained with Ponceau S (magenta color) as a control to ensure that the compounds did not induce nonspecific proteolysis (Lane M has MW markers). All compounds were run on the same gel. C The HNF4α level was quantified by ImageJ using the Western blots from panel B. Values represent the mean ± SE of 3 biological replicates, *p < 0.05, **p < 0.01 (vs scrambled siRNA or DMSO).

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