FIGURE 3.

RNF170 is an integral ER membrane protein. A, HeLa cells were cotransfected with cDNAs encoding the ER marker DsRed2-ER and wild-type RNF170^FLAG and were examined by confocal microscopy. B, HeLa cells transfected with cDNA encoding RNF170^FLAG were harvested in hypotonic buffer, sonicated, and fractionated by centrifugation (lanes 1 and 2). Pellets were then resuspended in hypotonic buffer, 0.1% SDS, or 0.1 m Na₂CO₃, and were recentrifuged (lanes 3–8). The supernatants (S) and pellets (P) from each centrifugation were then probed in immunoblots for the indicated proteins.