*4.6. Morphological Analysis: Hematoxylin and Eosin (H&E) Staining*

RAW 264.7 were seeded in Poly-D-lysine-coated glass dishes for 24 h then treated with LPS and *T. lutea* F&M-M36 extract, FX, or Celecoxib as described above. After 18 h, cells were fixed with 4% (w/v) paraformaldehyde for 15 min at room temperature. Next, cells were washed in H2O and then stained with hematoxylin for 2 min, differentiated in saturated lithium carbonate solution for 30 s, stained with eosin for 2 min, and dehydrated with ethanol series (50, 75, 96, and 100%), and finally xylene. Subsequently, glass dishes were mounted on microscope slides with a mounting medium and allowed to dry. Microscopic analysis was performed with ACT-2U software program (Nikon, Instruments Europe, Badhoevedorp, The Netherlands) connected via a camera to the microscope (Optiphot-2; Nikon). Five photomicrographs were randomly taken for each sample to evaluate cell morphology. The percentage of cells with dendritic changes (number of cells with clear morphological changes/total number of cells in the field × 100) were counted using ImageJ 1.33 image analysis software (http://rsb.info.nih.gov/ij (accessed on 22 April 2021)).
