Fig. 4.

AQP1 recruited ANXA2 to the Golgi apparatus and promoted the Golgi extension through F-actin by interaction with ANXA2, inducing breast cancer cell invasion. a Immunofluorescent assay showed the colocalization of ANXA2 and GM130 in Flag-vector/MDA-MB-231 and Flag-AQP1/MDA-MB-231 cells (n = 30 cells per group, two-tailed Student’s t test, ^***P < 0.001). Scale bar = 20 μm. b-c Immunofluorescent staining of GM130/TGN46 and DAPI in 4 different cell clones. In the right panel showed the distribution of cells with different ranges of Golgi ribbon angle (Ɵ: 0°–360°), respectively (n = 74–97 cells per group, ^***P < 0.001). Scale bar = 20 μm. d Flag-AQP1/MDA-MB-231 cells showed extended Golgi morphology (GM130, green) and normal F-actin (Texas red phalloidin). When treated with latrunculin B, Flag-AQP1/MDA-MB-231 cells exhibited loss of F-actin (stress fibers and peripheral actin) and Golgi condensation. Flag-AQP1/shANXA2 #1/MDA-MB-231 cells also reversed the extended Golgi morphology when treatment with latrunculin B. Scale bar = 20 μm. e The abilities of migration and invasion were detected using four indicated cells. Each bar represented the mean ± SEM from three or four independent experiments (two-tailed Student’s t test, ^*P < 0.05, ^**P < 0.01). Scale bar = 100 µm. All in vitro experiments were repeated at least 3 or 4 times