FIGURE 5:

Disruption of actin filaments leads to astral microtubule– and dynein-dependent cortical dissociation and spindle pole accumulation of LGN. (A) MDCK II cells were either untreated (Control) or treated as labeled. LatA: 1 μM of LatA for 45 min; Nocodazole + LatA: 50 nM of nocodazole plus 1 μM of LatA for 45 min; DYNC1H1 shRNA-1, -2 + LatA: transfected with DYNC1H1 shRNA-1 or -2 for 48 h and then treated with 1 μM LatA for 45 min. MG132, 5 μM, was added 1 h before treatments and maintained during treatments. Cells were fixed after treatments and stained with anti-LGN (green), anti–α-tubulin (red) antibodies, and DNA dye (blue). (B) Quantitation of LGN signals at spindle poles as described in Materials and Methods. n = 50 for each set; *p < 0.01.