Fig. 9. Syncollin-GFP is enriched in actin-coated structures in lacrimal acini exposed to BDM or ML-7. (A) Confocal micrographs of lacrimal acini transduced with Ad-Syncollin-GFP on day 2 of culture. On day 3 of culture, acini were stimulated with CCH (100 µM) for 5 minutes after pretreatment without (CON) or with BDM (10 mM) or ML-7 (40 µM) for 15 minutes. Cells were fixed with 4% paraformaldehyde to preserve Syncollin-GFP fluorescence (green) and labeled in parallel with rhodamine-phalloidin to visualize actin filaments (red). (B) Cells were treated and fixed as in A.; serial Z-stacks of images were obtained at 0.25 µm intervals and compiled into projection images in transparency mode using the 3D function of the Zeiss LSM software. Acinar thickness was 8-10 µm. Arrows, syncollin-GFP-enriched SVs beneath the actin-enriched lumen; arrowheads, syncollin-GFP-enriched SVs enveloped in actin coats; *, lumena; bars, 5 µm.