Three forms of autophagy are commonly described: macroautophagy, microautophagy, and chaperone-mediated autophagy (CMA). In macroautophagy, expendable cytoplasmic constituents are targeted and isolated from the rest of the cell within a double-membraned vesicle known as an autophagosome, which, in time, fuses with an available lysosome, bringing its specialty process of waste management and disposal; and eventually the contents of the vesicle (now called an autolysosome) are degraded and recycled. Chloroquine administration What is putative chloroquine transporter Plaquenil visual field 10-2 Inhibition of autophagic flux using chloroquine prevented clearance of p62 aggregates, leading to caspase-8 activation and cell death in C4-2 cells. In PC3 cells, inhibition of autophagy induction prevented p62 accumulation and hence caspase-8 activation. Abstract 3782 Chloroquine induces defective autophagic flux and enhances pancreatic cancer cell death induced by pegylated-arginine deiminase. Rouzbeh Daylami. Ing the basal autophagic flux by decreasing autophagosome-lysosome fusion, and not by inhibiting lysosomal degradation capacity as BafA 1 does. Although treatments with both CQ and BafA 1 ultimately cause a block of the autophagic flux, we show that the intracellular changes that are caused by these 2 compounds are profoundly different. In the extreme case of starvation, the breakdown of cellular components promotes cellular survival by maintaining cellular energy levels. In disease, autophagy has been seen as an adaptive response to stress, promoting survival of the cell; but in other cases it appears to promote cell death and morbidity. Autophagic flux chloroquine Restoration of Autophagic Flux Rescues Oxidative Damage and., Abstract 3782 Chloroquine induces defective autophagic flux. Sulfasalazine vs hydroxychloroquine Assays that monitor autophagic flux, or degradative completion of autophagy, are crucial for the assessment of the dynamic autophagy process in a variety of systems. Such assays help to distinguish between an increase in autophagosomes resulting from induced autophagic activity versus an increase in autophagosomes due to reduced lysosomal turnover. Monitoring Autophagic Flux by Using Lysosomal Inhibitors and Western.. Chloroquine inhibits autophagic flux by decreasing autophagosome.. Tenovin-6 impairs autophagy by inhibiting autophagic flux.. Autophagic flux assay in starved cells with a western blot of LC3. The correct way to calculate autophagic flux based on the LC3-II band is calculated below. Data and densitometric values are hypothetical. Novus Biologicals LC3B antibody was used in this example. Chloroquine is a lysosomotropic weak base, which in the monoprotonated form diffuses into the lysosome, where it becomes diprotonated and becomes trapped. Protonated chloroquine then changes the lysosomal pH, thereby inhibiting autophagic degradation in the lysosomes. Chloroquine is a known inhibitor of autophagic flux through the pathway, and chloroquine treatment results in an increase of LC3B puncta in otherwise normal cells. In contrast, siRNA-mediated knockdown of ATG7 expression results in a loss of LC3B puncta after chloroquine treatment Figure 3.