To quantify this observation, we randomly selected 12 PruΔgra12 and 10 PruΔku80 3-day-old in vitro cysts and measured the ratio of DBA fluorescence intensity at the cyst periphery, reflecting CST1 cargo delivered to the cyst wall, compared to the fluorescence intensity detected in the cyst interior, which highlights CST1 cargo not yet delivered to the cyst wall. Here, CST1 is linked to cyst.