Fig. 2

Assessment of proliferative activity in 3D canine colonoids exposed to palmitic acid. A Representative confocal microscopy images displaying 3D colonoids cultivated within Matrigel following a 24-h exposure to 30 µM of palmitic acid. Actively proliferating cells were marked by EdU staining (Cyan Blue), accompanied by nuclear counterstaining (DAPI; Blue). The left panel presents the image following 24 h of exposure to the carrier control (DMSO), while the right panel depicts the image after 24 h of exposure to 30 µM of palmitic acid. Scale bar = 25 µm. B The percentage of EdU-positive cells per nucleated cells in each condition was quantified using ImageJ. Each dataset was compiled from three biological replicates, each with three technical replicates. For each condition, five randomly chosen fields of view at 40 × magnification were selected from each sample, evaluating a minimum of ten organoids. The error bars represent the standard error of the mean. ** p < 0.01