To study the cellular localization of ICB, HT29 cells were seeded and grown to 70% confluency on 35 x 10 mm clear wall glass bottom dishes (Pelco, USA). The cells were then incubated with 5 μM ICB in media for either 2 hours or 24 hours at 37 °C and 5% CO2 in a humidified incubator. At the end of ICB incubation, cells were washed twice with 1XPBS and incubated with 1 μM MitoTracker Red or LysoTracker Red for 15 minutes at 37 °C and 5% CO2 in a humidified incubator. Next, the cells were washed twice with 1XPBS and fixed with 4% cold paraformaldehyde for 20 minutes at room temperature. After a single wash with 1X PBS, the cells were co-stained with 3 μM Hoechst 33342 for 10 minutes. After a single wash of 1X PBS, cells were imaged using Keyence BZ-X810 fluorescence microscope using a UV filter (ex: 387/11, em: 447/60), Cy5 filter (ex: 604/40, em: 672/50), and TxRd filter (ex: 559/34, em: 639/69). For each micrograph, a background subtraction with a rolling ball radius of 100 pixels was applied using Jackson lab BALB/c female mice weighing > 20 g was used for the procedure. A chemically induced paw inflammation model was established using the following steps developed based on the literature. Lipopolysaccharide (LPS) from E. coli was used as an inflammation stimulator.