Journal: Frontiers in Microbiology

Article Title: Replication of Dengue Virus in K562-Megakaryocytes Induces Suppression in the Accumulation of Reactive Oxygen Species

doi: 10.3389/fmicb.2021.784070

Figure Lengend Snippet: DENV replication suppresses ROS in differentiating cells by upregulation of NFE2L2 activity. (A) K562 cells either uninfected or infected with DENV at an MOI of 1.0 were differentiated with 50 nM PMA in the absence or presence of 10 μM of RA839. At 3 or 6 days post-infection the cytosolic ROS was quantified by H 2 DCFDA staining as described above. (B) K562 cells infected with DENV at an MOI of 1.0 were differentiated with PMA in the absence or presence of 10 μM of RA839. The supernatant was collected at 3 or 6 days post-infection and the infectious titer of secreted virus quantified by Focus-forming unit (FFU) assay. The Log 10 of the FFU/ml was calculated and plotted. (C) K562 cells either uninfected or infected with DENV at an MOI of 1.0 were differentiated with PMA in the absence or presence of 10 μM of ML385. At 3 or 6 days post-infection the cytosolic ROS was quantified by H 2 DCFDA staining as described above. (D) K562 cells infected with DENV at an MOI of 1.0 were differentiated with PMA in the absence or presence of 10 μM of ML385. The culture supernatant was collected at 3 or 6 days post-infection and the infectious titer of secreted virus quantified by Focus-forming unit (FFU) assay. The Log 10 of the FFU/ml was calculated and plotted. (A–D) The error bars represent standard deviation obtained from 3 independent experiments and the P -values of significance calculated by Student's t -test are indicated.

Article Snippet: Phorbol-12 Myristate-13 acetate (PMA, Sigma Aldrich), N-Acetyl Cysteine (NAC, Sigma Aldrich), NITD008 (Sigma Aldrich), RA839 (Tocris), ML385 and Sodium Butyrate (Sigma Aldrich) were diluted in recommended vehicles and stored as single use aliquots at −20°C.

Techniques: Activity Assay, Infection, Staining, Virus, Standard Deviation