Confocal immunofluorescent analysis of He La (upper) and C2C12 (lower) cells, chloroquine-treated (50 μM, overnight; left), nutrient-starved with EBSS (3 hr, middle) or untreated (right) using LC3A/B (D3U4C) XP Immunohistochemical analysis of paraffin-embedded human squamous cell lung carcinoma using LC3A/B (D3U4C) XP® Rabbit m Ab in the presence of control peptide (left) or antigen-specific peptide (right). Western blot analysis of extracts from He La cells, mock transfected or transfected with rat LC3B, and from HT-1080 and A20 cells, untreated or chloroquine-treated (50 μM, overnight), using LC3B Antibody. Chloroquine synthesis reaction Hydroxychloroquine shingles Chloroquine CQ has been under clinical use for several decades, and yet little is known about CQ sensing and signaling mechanisms or about their impact on various biological pathways. We employed the budding yeast Saccharomyces cerevisiae as a model organism to study the pathways targeted by CQ. Chloroquine is a lysosomotropic agent that prevents endosomal acidification 1. It accumulates inside the acidic parts of the cell, including endosomes and lysosomes. This accumulation leads to inhibition of lysosomal enzymes that require an acidic pH, and prevents fusion of endosomes and lysosomes. When trafficking and/or acidification is disrupted by chloroquine or bafilomycin A1, Toll-like receptor signaling is inhibited 1, 14, 30, 32, 37. In addition to these studies in vitro, a few reports suggest that chloroquine can inhibit innate immune responses in vivo in a 2-hit model of hemorrhage then CLP 13, after CpG/LPS administration 18, and in a mouse cryptococcosis infection model 31. Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye). Confocal immunofluorescent analysis of HCT-116 cells, untreated (left) or choroquine-treated (50 u M, overnight; right) using LC3B Antibody (green) and β-Catenin (L54E2) Mouse m Ab (Alexa Fluor® 555 Conjugate) #5612 (red). Chloroquine cell signalling Cell Signaling Technology, Chloroquine for research Cell-culture tested InvivoGen What does the drug plaquenil do When imaging using a confocal microscope there is no clear LC3 signal, and definitely no LC3 positive punctae as you would expect to see. I treated some cell lines with 25 mM Chloroquine for. Questions with answers in CHLOROQUINE Science topic. Chloroquine and inhibition of Toll-like receptor 9 protect.. Chloroquine - Wikipedia. Chloroquine binds to heme or FP to form what is known as the FP-Chloroquine complex; this complex is highly toxic to the cell and disrupts membrane function. Action of the toxic FP-Chloroquine and FP results in cell lysis and ultimately parasite cell autodigestion. In essence, the parasite cell drowns in its own metabolic products. Incubate membrane and primary antibody at the appropriate dilution and diluent as recommended in the product datasheet in 10 ml primary antibody dilution buffer with gentle agitation overnight at 4°C. Wash three times for 5 min each with 15 ml of TBST. Hydroxychloroquine and chloroquine are weak bases and have a characteristic ‘deep’ volume of distribution and a half-life of around 50 days. such as TLR7 and TLR9 signalling, T cell.