Pex30 complexes contribute to MCS formation. (A) Single Z-slices of WT spheroplasts during the exponential phase and diauxic shift from volumes acquired using SBF-SEM 3View. The site of the minimal distance between the nucleus and the vacuole is indicated by an arrowhead. N, nucleus; V, vacuole; LD, lipid droplet; SBF-SEM, serial block-face scanning electron microscopy. Bars, 1 µm. (B) Single Z-slices (left) and volume reconstruction (right) of WT, pex30Δ, pex29Δ, and nvj1Δ spheroplasts during the diauxic shift acquired using FIB-SEM. The ER is reconstructed in green, the vacuole in magenta, and LDs in yellow. Bar, 1 µm. (C) Localization of endogenous Pex3-mNG and Inp1-mCherry in exponential growing WT cells. Bar, 5 µm. (D) Quantification of Pex3 and Inp1 protein levels in cells in the indicated genotype. Three independent experiments were conducted, and Student’s t test (two-tailed) was performed to compare the normalized intensity between conditions (ns, P > 0.5). Bars represent the SD. FIB-SEM, focused ion beam scanning electron microscopy.