Outline and workflow of the high-content screening assay. (a) Schematic representation of NRK49F cells towards fibrosis and high-content screening (HCS) assay outline. NRK49F cells were seeded in 96-well plates at a density of 6,000 cells per well. Compounds were added, and after 72 h of culture, cells were fixed, stained, and subjected to high-content imaging analysis using MetaXpress Micro XL. A schematic approach for systematic identification of compounds with fibrosis inhibiting potential. Identification of the hit compounds was based on the above images using high-content analysis. The final selected compound from the screen included Hydroxysafflor Yellow A, Crocin, and Gypenosides. (b) Workflow of the study.