Golgi ribbon reassembly is impaired in PAFAH Ib knockdown cells. (A) Confocal images of the Golgi (anti-ManII) before BFA, after 20 min in BFA (0 min), and 20, 45, or 90 min after BFA washout in control or α1 and α2 siRNA-transfected cells. (B) Percentage of cells with reassembled Golgi ribbons after BFA washout. n = 4; error bars = SEM; at 90 min P < 0.01 by t test. (C) Wide-field fluorescence images of the Golgi (anti-ManII) before BFA treatment, and after 0, 20, 45, or 60 min of BFA washout. RNAi-resistant HA-LIS1 expression is shown in insets (anti-HA). (D) Percentage of cells with reassembled Golgi ribbons; n = 3; error bars = SEM. (E) Golgi mini-stacks (anti-ManII) after 30 min of BFA recovery for α1 and α2 siRNA or LIS1 siRNA, or α1/α2/LIS1 siRNA (triple)-treated BTRD cells. LIS1 knockdown cells show extensive membrane tubules, which are not seen in α1 and α2 or triple knockdown cells. (F) Percentage of cells with Golgi tubules after 30 min of BFA washout. Bars: (A and C) 10 µm; (E) 5 µm.