Journal: The FASEB Journal

Article Title: Atorvastatin restricts the ability of influenza virus to generate lipid droplets and severely suppresses the replication of the virus

doi: 10.1096/fj.201900428RR

Figure Lengend Snippet: Autophagy increases lipid droplets and is regulated by ER stress during infection. A) Autophagy inhibitor 3MA pretreatment at 5 mM, applied 1 h before infection, partially blocks the induction of lipid droplets by IAV (IAV+3MA vs. IAV). B) This reduction during autophagy inhibition is significant (P < 0.05). Complete field shown. Original magnification in both A and B, ×400. C) Viral replication persists in the presence of 3MA. D) Influenza-induced autophagy is inhibited by ER stress inhibitor salubrinal (Sal), suggesting ER stress regulation of influenza induced autophagy. At least 3 experiments were run for each condition. Blue = DAPI; red = ORO; green = viral NP. Triple fluorescence experiments are not shown because of technical limitations, though correspondence of lipid droplets with infection is illustrated in Fig. 5. βtub, β-tubulin. ** P<0.01, *** P<0.001.

Article Snippet: When appropriate, cells were treated with class III PI3K inhibitor 3-methyladenine (3MA) (M9281; MilliporeSigma) at 5 mM ( 38 ) ( 3 , 39 ), salubrinal (sc-202332; Santa Cruz Biotechnology, Dallas, TX, USA) at 5 μM ( 40 ), N -acetyl cysteine (NAC) at 5 mM (ENZ-51010; Enzo Life Sciences, Farmingdale, NY, USA) ( 41 , 42 , 43 ).

Techniques: Infection, Inhibition, Fluorescence

Journal: The FASEB Journal

Article Title: Atorvastatin restricts the ability of influenza virus to generate lipid droplets and severely suppresses the replication of the virus

doi: 10.1096/fj.201900428RR

Figure Lengend Snippet: A model for the site of action for the ability of ATV to suppress replication of influenza virus. Infection stimulates ER stress, which through increased production of ROS and other mechanisms up-regulates autophagy. Increased autophagy and other presumptively metabolic changes induced by infection ultimately activate HMG-CoA reductase, which is required for the production of lipid droplets and is blocked by ATV. In the absence of ATV, production of ER stress signals and ROS and up-regulation of autophagy can be individually inhibited but still allow alternative pathways to the synthesis of lipids in infected cells. Individual blockage of any one of these metabolic processes fails to prevent synthesis of viral protein and presumptive production of virus. When HMG-CoA reductase is inhibited by ATV, production of lipid droplets and replication of virus are blocked. Under the conditions we used, the combination of 3MA and salubrinal (SAL) is too toxic to evaluate for reliable results.

Article Snippet: When appropriate, cells were treated with class III PI3K inhibitor 3-methyladenine (3MA) (M9281; MilliporeSigma) at 5 mM ( 38 ) ( 3 , 39 ), salubrinal (sc-202332; Santa Cruz Biotechnology, Dallas, TX, USA) at 5 μM ( 40 ), N -acetyl cysteine (NAC) at 5 mM (ENZ-51010; Enzo Life Sciences, Farmingdale, NY, USA) ( 41 , 42 , 43 ).

Techniques: Virus, Infection