cell culture imr 90  (ATCC)

Journal: PLOS Pathogens

Article Title: NS5-targeting nucleoside analogs inhibit dengue virus and other flaviviruses

doi: 10.1371/journal.ppat.1013970

Figure Lengend Snippet: Representative fluorescence microscopy images of DENV2 infection in the indicated cell models (A549, IMR90, HepG2, and Huh7.5).Cells were treated with vehicle or the indicated compounds and either left uninfected or infected with DENV2 at MOI 2 (Huh7.5 and A549), MOI 1 (HepG2), and MOI 5 (IMR90). At 24 hours post-infection (hpi), cells were fixed and stained for viral antigen (4G2, green) and nuclei (Hoechst 33342, blue). Images were acquired at 10 × magnification. B. Schematic of the screening strategy. 384-well format high-throughput screen was conducted using A549, IMR90, HepG2, and Huh7.5 cells to evaluate a library of 1,101 nucleoside analogs for their ability to inhibit DENV2 infection at 50 μM. Cells were treated with compounds for 2 h then infected with DENV2 at the MOI in A, followed by fixation at 24 hours post-infection. Automated microscopy was used to quantify percent of infection and cell numbers normalized to vehicle control. Percent of control (POC) for DENV2 infection in C. Huh7.5 D. HepG2 cells E. A549 cells and F. IMR90 cells. G. Venn diagram representing 68 non-redundant candidates that showed >80 inhibition of infection; > 60% viability in at least one of the screened cell models (C-F) . H. Summary of hit selection and downstream validation showing the progression from 68 initial hits identified in four cell models to 23 candidates with SI > 5 based on dose–response analysis, followed by qPCR validation in selected cell models.

Article Snippet: IMR90 cells (CCL-186) were obtained from ATCC and cultured in DMEM-20 (4.5g/L glucose with Na-pyruvate) supplemented with 20% (v/v) fetal bovine serum and were maintained at 37°C and 5% CO 2 .

Techniques: Fluorescence, Microscopy, Infection, Staining, High Throughput Screening Assay, Control, Inhibition, Selection, Biomarker Discovery