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Additional Supporting Information may be found in the online version of this article.

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dvdy24000-sup-0001-suppmovie1.mpg1312KSupp. Movie S1. Time-lapse movie of salivary gland branching morphogenesis. Mouse embryonic day (E) 13.5 submandibular salivary gland was maintained in three-dimensional organ culture and imaged by brightfield microscopy for 96 hr.
dvdy24000-sup-0002-suppmovie2.AVI6532KSupp. Movie S2. Immunofluorescence time-lapse movie of photo-converted outer bud cells. This movie shows images acquired using a spinning disk confocal microscope every 10 min in Z-stacks and presented as 20 μm maximum intensity projections. For this and all subsequent movies, frame rate is 15 fps for images acquired at a rate of 1 frame/10 min. The initial photo-conversion highlighted a single epithelial region containing at least 12–16 cells. Left: red channel; center: green channel; right: overlay of red and green.
dvdy24000-sup-0003-suppmovie3.AVI1452KSupp. Movie S3. Outer bud cell leaving the basement membrane to undergo cell division. This fluorescence time-lapse movie shows a single outer bud cell migrating away from the basement membrane and then undergoing mitosis; one of the daughter cells returns to the basement membrane. Images were acquired every 10 min and presented as described for Movie 2. Left: red; center: green; right: overlay.
dvdy24000-sup-0004-suppmovie4.AVI14449KSupp. Movie S4. Inner bud cells displaying lower motility. This fluorescence time-lapse movie acquired under the same conditions as the other movies shows slower average motility of cells photo-converted in a region of the inner bud. Left: red; center: green; right: overlay.
dvdy24000-sup-0005-suppmovie5.AVI7221KSupp. Movie S5. Outer bud cells in a salivary gland treated with antibodies against α6 and β1 integrin antibodies at 100 μg/ml each. Note the loss of association of the migrating cells with the basement membrane. Left: red; center: green; right: overlay.
dvdy24000-sup-0006-suppmovie6.AVI11693KSupp. Movie S6. Composite movie showing effects of inhibiting integrins on outer bud cell motility rates and patterns. The left panel shows untreated control inner bud cells, and the right panel shows inner bud cells treated with 100 μg/ml inhibitory anti-α6 and -β1 integrin antibodies (“Integrin I.”). Both panels show overlay of red and green channels. Note the loss of cell association with the basement membrane after integrin inhibition, as well as the decreased average velocity of migration of the cells, reflected in the reduced lateral dispersion of cells along the basement membrane.
dvdy24000-sup-0007-suppmovie7.AVI7651KSupp. Movie S7. Outer bud cells in a salivary gland treated with inhibitory anti-E-cadherin monoclonal antibody (100 μg/ml ECCD-1). Note the continued association of some of the cells with the basement membrane. Left: red; center: green; right: overlay.
dvdy24000-sup-0008-suppmovie8.AVI21278KSupp. Movie S8. Composite movie showing effects of inhibiting E-cadherin on inner and outer bud cells of salivary glands. The upper panels show untreated control inner and outer bud cells, and the lower panels show these cells in glands treated with inhibitory anti-E-cadherin monoclonal antibody (100 μg/ml ECCD-1, labeled Ecad). For clearer visualization of the red photo-converted cells, the left panels show only the red channel in grayscale, while the right panels show overlay of the red and green channels. Note the disruption of cell–cell adhesion and the formation of black gaps or holes in the inner bud epithelium.
dvdy24000-sup-0009-suppmovie9.AVI3527KSupp. Movie S9. Outer bud cells in a salivary gland treated with 50 μM blebbistatin. Note the continued association of some of the cells with the basement membrane. Left: basement membrane stained for collagen IV; center: photo-converted outer bud cells; right: overlay of collagen IV (pseudo-colored green) and red channel showing photo-converted KikGR outer bud cells. The collagen IV was imaged using a 642-nm laser, because the green channel cannot be used to avoid blebbistatin photo-inactivation and resultant cytotoxicity.
dvdy24000-sup-0010-suppmovie10.AVI7869KSupp. Movie S10. Composite movie comparing control salivary gland with glands treated with monoclonal antibody inhibitors of E-cadherin (Ecad) or integrin α6 and β1 subunits (Integrin), or with blebbistatin to inhibit myosin II isoforms (Myosin II). For all four panels, red indicates photo-converted cells, and green indicates non–photo-converted KikGR-expressing cells in panels A–C. In panel D, the basement membrane visualized using a collagen IV antibody imaged at 642 nm and pseudo-colored green to avoid blebbistatin degradation and cytotoxicity.

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