Figure 4.

Silencing of RINT-1 causes dispersion of proteins located in compartments of the early secretory pathway. (A) HeLa cells were transfected without (mock) or with lamin A/C siRNA, RINT-1 (268), or RINT-1 (1149). At 72 h after transfection, the cells were solubilized in phosphate-buffered saline with 0.5% SDS. Equal amounts of total proteins were separated by SDS-PAGE and analyzed by immunoblotting with the indicated antibodies. (B) Immunofluorescence microscopic analysis of mock-treated cells, and cells transfected with lamin A/C siRNA, RINT-1 (268), or RINT-1 (1149). At 72 h after transfection, the cells were fixed and double-stained with antibodies against RINT-1 and p115. Expanded images for p115 are shown on the right. Bar, 5 μm. (C) Cells were treated as described in B and double-stained with antibodies against Sec31p and ERGIC-53. Bar, 5 μm.