Figure 7.

CHC knockdown minimally affects Golgi reformation in response to nocodazole treatment. NRK cells were transfected with either control nonsilencing siRNA or with CHC siRNA (rchc2) for 3 d, after which they were untreated (a–e) or treated with 30 μM nocodazole for 90 min (f–j). Nocodazole treatment triggered GRASP65 redistribution into peripherally distributed immunoreactive puncta in cells with nonsilencing or rchc2 siRNA (g and i). The medium was replaced (k–o), after which clathrin and GRASP65 were localized using mouse anti-CHC antibody (X22) and rabbit anti-GRASP65 antibody. In rchc2-transfected cells, GRASP65 regained its normal morphology less efficiently than in untransfected cells (n; asterisk) or cells transfected with nonsilencing siRNA (l). Asterisk indicates siRNA-transfected cells. Bar, 20 μm.