Journal: PLoS ONE

Article Title: Fomiroid A, a Novel Compound from the Mushroom Fomitopsis nigra , Inhibits NPC1L1-Mediated Cholesterol Uptake via a Mode of Action Distinct from That of Ezetimibe

doi: 10.1371/journal.pone.0116162

Figure Lengend Snippet: A, HEK293 cells stably expressing hNPC1L1-EGFP (WT) or hNPC1L1 L1072T/L1168I -EGFP (L1072T/L1168I) were incubated in the absence or presence of 10 µM fomiroid A for 24 h, and then plasma membranes (PM) were stained with CellMask Orange. The arrows show the rescue of mislocalized L1072T/L1168I mutant by fomiroid A treatment. Scale bar, 10 µm. B, WT or L1072T/L1168I mutant cells were treated with the indicated concentrations of fomiroid A for 24 h. The amount of NPC1L1 was determined using anti-GFP antibody. Vinculin was used as a loading control. C, Intensities of bands representing mature NPC1L1. Values represent means ± S.E. (n = 3). * p <0.05 compared with untreated mutant cells.

Article Snippet: The following antibodies were used in this study: mouse monoclonal anti-HA (F-7, sc-7392, Santa Cruz Biotechnology), anti-GFP (B-2, sc-9996, Santa Cruz Biotechnology), anti-vinculin (V9131, Sigma-Aldrich), rabbit polyclonal anti-NPC1L1 (HPA018105, Sigma-Aldrich), horseradish peroxidase (HRP)-rabbit anti–mouse IgG (H+L) conjugate (81-6720, Life Technologies), HRP-goat anti-rabbit IgG (H+L) conjugate (81-6120, Life Technologies), and Alexa Fluor 633 goat anti–mouse IgG (H+L) conjugate (A-21052, Life Technologies).

Techniques: Stable Transfection, Expressing, Incubation, Staining, Mutagenesis

Journal: PLoS ONE

Article Title: Fomiroid A, a Novel Compound from the Mushroom Fomitopsis nigra , Inhibits NPC1L1-Mediated Cholesterol Uptake via a Mode of Action Distinct from That of Ezetimibe

doi: 10.1371/journal.pone.0116162

Figure Lengend Snippet: A, Expression of NPC1L1 was determined by western-blot analysis using anti-NPC1L1 antibody. B, Differentiated Caco2/mock and Caco2/rNPC1L1 cells were incubated for 1 h at 37°C with a micellar solution containing 2 mM sodium taurocholate, 50 µM phosphatidylcholine, 1 µM cholesterol, 1 µCi/ml [ 3 H]cholesterol, and the indicated concentrations of ezetimibe or fomiroid A. Radioactivity of [ 3 H]cholesterol was counted in a liquid scintillation counter. C, Differentiated Caco2/mock and Caco2/rNPC1L1 cells were incubated at 37°C with a micellar solution containing 5 mM sodium taurocholate, 500 µM oleate, 10 µM cholesterol, 1 µCi/ml [ 3 H]cholesterol, and the indicated concentrations of ezetimibe or fomiroid A; after 1 h, the micellar solution was replaced with DMEM containing Insulin-Transferrin-Selenium. The cells were incubated for 8 h at 37°C. The lipids extracted by organic solvent were separated with TLC, and the radioactivity of [ 3 H]esterified cholesterol was counted by a liquid scintillation counter. Values represent the means ± S.E. (n = 3). * p <0.05, ** p <0.01 compared with control Caco2/mock cells, # p <0.05, ## p <0.01 compared with control Caco2/rNPC1L1 cells.

Article Snippet: The following antibodies were used in this study: mouse monoclonal anti-HA (F-7, sc-7392, Santa Cruz Biotechnology), anti-GFP (B-2, sc-9996, Santa Cruz Biotechnology), anti-vinculin (V9131, Sigma-Aldrich), rabbit polyclonal anti-NPC1L1 (HPA018105, Sigma-Aldrich), horseradish peroxidase (HRP)-rabbit anti–mouse IgG (H+L) conjugate (81-6720, Life Technologies), HRP-goat anti-rabbit IgG (H+L) conjugate (81-6120, Life Technologies), and Alexa Fluor 633 goat anti–mouse IgG (H+L) conjugate (A-21052, Life Technologies).

Techniques: Expressing, Western Blot, Incubation, Radioactivity

Journal: Pharmaceuticals

Article Title: High-Efficacy α,β-Dehydromonacolin S Improves Hepatic Steatosis and Suppresses Gluconeogenesis Pathway in High-Fat Diet-Induced Obese Rats

doi: 10.3390/ph14040375

Figure Lengend Snippet: α,β-dehydromonacolin S ( C5 ) modulates lipid transporter gene and protein expression. ( a ) The mRNA expression of lipid transporters including CD36, NPC1L1, LDLR, ABCG5/8 and ABCA1 were quantitatively determined using qPCR. ( b ) CD36 and ( c ) NPC1L1 protein expression extracted from rat liver tissue were analyzed by Western blotting. A representative blot of CD36 and NPC1L1 protein expression is shown on the top panel and quantification of relative protein expression in each experimental group presented on the bottom. Anti-β-actin antibody was used as loading control. Values are mean ± S.E.M ( n = 5). * p < 0.05 represents the significant difference compared with the NDV group and # p < 0.05 represents the significant difference compared with the HFV group.

Article Snippet: Polyclonal rabbit anti-Niemann–Pick C1-Like 1 (NPC1L1) and cluster of differentiation 36 (CD36) were purchased from Novus biological (CO, USA).

Techniques: Expressing, Western Blot, Control

Journal: FASEB bioAdvances

Article Title: Identification of hepatic NPC1L1 as an NAFLD risk factor evidenced by ezetimibe‐mediated steatosis prevention and recovery

doi: 10.1096/fba.2018-00044

Figure Lengend Snippet: Key resources

Article Snippet: Rabbit polyclonal anti‐NPC1L1 , Novus Biologicals , Cat# NB400‐128; RRID: AB_10000815.

Techniques: Virus, Liposomes, Control, Microarray, Expressing, Recombinant