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Image Search Results
Journal: PLoS ONE
Article Title: Inhibitory effects of fenretinide metabolites N-[4-methoxyphenyl]retinamide (MPR) and 4-oxo-N-(4-hydroxyphenyl)retinamide (3-keto-HPR) on fenretinide molecular targets β-carotene oxygenase 1, stearoyl-CoA desaturase 1 and dihydroceramide Δ4-desaturase 1
doi: 10.1371/journal.pone.0176487
Figure Lengend Snippet: A) SCD1 protein expression is decreased after treatment with fenretinide, and its metabolite 3-keto-HPR. ARPE-19 cells were treated with the indicated concentration of fenretinide, its metabolites, A1120, or vehicle for 24 h. Cell lysates were analyzed by Western blotting using mouse monoclonal antibody against SCD1 (upper panel) in comparison with immunoreactivity to α-tubulin which was employed as a loading control (lower panel) and densitometry for each lane was performed and analyzed by ImageJ to provide the SCD1/α-tubulin ratio for each lane. B) SCD1 protein activity is decreased after treatment with fenretinide, and its metabolite 3-keto-HPR. ARPE-19 cells were treated with the indicated concentration of fenretinide, its metabolites, A1120, or vehicle for 16 h, then incubated an additional 5 h in the presence of methyl D3-palmitic acid (50 μM) before harvesting. Free fatty acids present in lipid extracts were analyzed by LC/MS and SCD1 activity, measured by the conversion of methyl-D3 palmitic acid to methyl-D3 palmitoleic acid, was determined. Fenretinide and 3-keto-HPR are effective inhibitors of SCD1 activity, whereas MPR and A1120 have no inhibitory effect on SCD1 activity at all. Lipids were extracted from two 100 mm Petri dishes and combined for mass spectrometry analysis. Data plotted as means ± standard deviation; n = 3.
Article Snippet: After blocking in 5% non-fat milk in Tris-buffered saline (TBS) containing 0.01% Tween 20 for 1 h, the membranes were incubated overnight at 4°C with
Techniques: Expressing, Concentration Assay, Western Blot, Activity Assay, Incubation, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry, Standard Deviation