npc1 sc 271334 antibodies  (Santa Cruz Biotechnology)

Journal: Viruses

Article Title: Bafilomycin A1 Inhibits HIV-1 Infection by Disrupting Lysosomal Cholesterol Transport

doi: 10.3390/v16091374

Figure Lengend Snippet: NPC1 overexpression partially relieves bafilomycin A1 inhibition of HIV-1 infection. ( A ) TZM-bl cells were transfected for 48 h with a plasmid expressing NPC1-GFP or the empty control plasmid pcDNA3.1. Western blotting was conducted to confirm the expression of NPC1-GFP using an antibody specific for GFP or NPC1. GAPDH was included as a loading control. ( B ) Four hours after transfection with the NPC1-GFP expressing plasmid or the empty vector control, cells were treated with bafilomycin A1 (5 nM) for 48 h and free cholesterol was detected using filipin. Images (60×) were obtained using the Nikon A1R confocal microscope. Scale bar = 20 µm. Images are representative of three independent experiments. ( C ) At 24 h post-transfection with the NPC1-GFP expressing plasmid or the empty vector control, cells were infected with HIV-1 in the absence or presence of bafilomycin A1 (5 nM). At 48 h post-infection, HIV-1 infection was determined using a luciferase assay. Data are means ± SD ( n = 3). * p < 0.05. *** p < 0.001.

Article Snippet: Antibodies for GFP (sc-9996) and NPC1 (sc-271335) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

Techniques: Over Expression, Inhibition, Infection, Transfection, Plasmid Preparation, Expressing, Control, Western Blot, Microscopy, Luciferase