MLF2 localization correlates with K48-Ub enrichment at the nuclear periphery. (A) Confocal microscopy images demonstrating the colocalization between MLF2-HA and K48-Ub at the nuclear periphery of 4TorKO cells. Scale bar, 10 µm; inset, 2 µm. (B) Line-scan profiles of K48-Ub signal (green) and MLF2-HA signal (red) at the nuclear periphery of the cell from A. Measurements were taken for the region represented in the inset. (C) Graphical representation illustrating the percent of 4TorKO cells transfected with MLF2-HA that display K48-Ub foci or MLF2-HA foci at the nuclear periphery. Data shown are the mean of three independent experiments with 75 cells each, and error bars indicate ±SD. (D) Torsin-deficient cells were analyzed by immunoelectron microscopy to visualize the subcellular localization of Mab414 relative to endogenous MLF2 (upper panel), and transiently transfected MLF2-GFP (bottom panel) in blebs. Scale bars, 250 nm (top panel) and 500 nm (bottom panel). N, nucleus; C, cytoplasm.