The anti-mouse RetSat monoclonal antibody (red) was used to stain Tet-induced HEKK-RetSat-transfected cells (A) and untransfected cells (B). HEKK-RetSat cells stained with the anti-RetSat monoclonal antibody examined under higher magnification show the perinuclear and reticular membrane localization of RetSat in transfected cells (C). Scale bar, 20µm. D, subcellular analysis of RetSat protein in mouse liver cells. Immunoblotting of equal amounts of protein from the cytosolic supernatant, postnuclear membrane fraction, and whole cell lysate of mouse liver cells indicates that the RetSat protein is membrane-associated. An immunoreactive band of a protein with apparent molecular mass of 70 kDa was identified as the mouse RetSat protein, confirmed by its presence in the lysate of Tet-induced HEKK-RetSat cells. The blots were probed with the anti-mouse RetSat monoclonal antibody.