Figure 7

SSW smoke affects the endomembrane network. Cells were exposed to 4 or 8 hours of smoke treatment (SSW or MSW) and prepared for fluorescence imaging. (A) Confocal microscopy analysis of DIOC6 stained cells. Untreated cells show normal ER network concentrated around the nucleus and also spread out to the periphery of the cell. (B) SSW treated cells show that the ER is fragmented when compared with the control and is only found around the nucleus. (C) MSW-treated cells show similar ER morphology to that of the control. (D-F) To ascertain the status of the Golgi network, we used anti-β-COP monoclonal antibody for immunocytochemistry, which specifically labels the Golgi network. (D) In untreated cells, the staining of the Golgi surrounds the nucleus and vesicles are seen all over the cytosol. (E) In SSW-treated cultures, the cells have many fewer Golgi vesicle staining. (F) MSW treated cells show an organization similar to that found in the control. (G-I) Fibroblasts were treated as above and immunolabeled for cIL-8. (G) In untreated cells, the expression of cIL-8 is barely present because this protein is only expressed when it is stress-induced. (H) SSW-treated cells were observed to have no perinuclear staining; instead, the endomembrane system was dispersed throughout the cytosol. (I) MSW-treated cells show that the cIL-8 is being produced in the endomembrane system near the nucleus where this normally occurs. Pictures are representative of 2 different experiments. Scale bars = 50 μm in (A-F) and 30 μm in (G-I).