(A) Western blotting analysis confirmed a partial level of Rab6 depletion consisting of a drop of 40% in siRNA Rab6 macrophages, and (B) we also confirmed previous reports of collapsed, and frequently fragmented, Golgi complex compared with the most classical perinuclear shape distribution in control cells, using anti-GM130 as a Golgi marker. (C) Morphological changes on the Golgi complex in siRNA Rab6 macrophages LPS-activated for 2 h were visualized by EM; in control cells, the Golgi complex is formed by a multiple interconnected stacks (a, white arrowheads) approaching one to each other to form a ribbon. In siRNA Rab6 (b, white arrowheads) the Golgi complex consists of a huge single isolate ribbon, ticker and at least three times longer (white arrowheads), as previously characterized . The addition of LPS did not significantly change the Golgi morphology (c, white arrowheads). Original magnification has been reported in each image (and relative inset), and the reference bars present on large images (vertically, top right). (D) After 2 h, LPS-activated RAW 264.7 cells clearly present internal and plasma membrane (surface) TNF staining (asterisks), while in siRNA Rab6 cells the TNF remains trapped in the cell. Original optical magnification 63X (B, D). Bars: 5 µm (B), 1 µm (C), 10 µm (D). N, nucleus; m, mitochondrion; ER, endoplasmic reticulum.