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. Author manuscript; available in PMC: 2014 Aug 16.
Published in final edited form as: ACS Chem Biol. 2013 May 29;8(8):1730–1736. doi: 10.1021/cb4000986

Figure 5. Alverine and Benfluorex altered proteolytic sensitivity in HNF4α protein.

Figure 5

A) For the DARTS assay, HepG2 cells were treated with DMSO (lane 1), BI6018 (lane 2), BI6015 (lane 3), Alverine (lane 4) or Benfluorex (lane 5) at a concentration of 20µM or 40µM for 16hr. Total cell protein was extracted and each sample was split into two aliquots for proteolysis without (−) or with (+) subtilisin (A, left panels) and blotted with HNF4α antibody. After detection of HNF4α, the membrane was stained with Ponceau S (A, right panels) as a control to ensure that the compounds did not induce nonspecific proteolysis (Lane M-MW markers). All compounds were run on the same gel. A lane from a compound unrelated to the studies reported here was deleted, accounting for the gap between lanes 3 and 4. B) Western blots were quantified using ImageJ software, demonstrating a statistically significant effect of BI6015, alverine, benfluorex on subtilisin sensitivity. Values represent the mean ± SE of 3 biological replicates, *p<0.05.