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

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DVDY21709SuppFigS1.tif230KFig. S1. Live salivary gland lumen length and strain. (A–C) Measurements of WT and <I>rib</I> SG lumen length from live imaging. Movies are aligned with VM contact at t = 30 minutes. Rate of lumen length increase is similar within genotypes for WT (A) and <I>rib</I> (B) and is remarkably linear. Average <I>rib</I> SG lumen length is 60% of average WT SG lumen length at t = 100 minutes (C). Calculated lumen strain (based on starting length at t = 0 minutes) indicates that <I>rib</I> SGs achieve around 34% of WT strain during this time (D). Linear trendlines for average lumen length (C) are WT: y = 0.6635x + 26.341, R<SUP>2</SUP> = 0.9799 and <I>rib</I>: y = 0.2339x + 35.262, R<SUP>2</SUP> = 0.9740. Linear trendlines for average lumen strain (D) are WT: y = 0.0202x; R<SUP>2</SUP> = 0.9699 and <I>rib</I>: y = 0.0087x; R<SUP>2</SUP> = 0.9234. These trendlines indicate that <I>rib</I>salivary gland lumena increase in length at approximately 35% of the WT velocity and 43% of the WT strain rate.
DVDY21709SuppFigS2.tif2231KFig. S2. The effects of overexpression of wt Rib or non-phosphorylatable (dominant-negative) Moe. WT stage 12 and 14 SGs have narrow uniform lumena (arrows in A, D), whereas stage 12 and 14 SGs overexpressing either Rib (B, E) or Moe<SUP>T559A</SUP> (C,F), have wider, more irregular lumena (arrows in B, C, E, F). An antibody to Sas marks the SG lumena in black.
DVDY21709SuppFigS3.tif760KFig. S3. Finite element modeling of tube elongation (0.25x-2.50x of WT value). Simulations of tube elongation using elastic material model. Deformation and local strain plots for values of apical stiffness from 0.25x to 2.5x are shown. Keys for deformation and strain are at bottom of figure.
DVDY21709SuppFigS4.tif734KFig. S4. Finite element modeling of tube elongation (2.75x-5.00x of WT value). Simulations of tube elongation using elastic material model. Deformation and local strain plots for values of apical stiffness from 2.75x to 5x are shown. Keys for deformation and strain are at bottom of figure.
DVDY21709SuppFigS5.tif7845KFig. S5. Correction collar adjustment for spherical aberration. (A-J) Images of migrating salivary gland expressing α-catenin-GFP and DsRed<SUP>N</SUP> using the custom multiphoton microscope schematized in Figure 7 and an Olympus 60x 1.2 N.A. lens. Correction collar setting is denoted in the lower right. Adjustment of the correction collar from the standard setting of 0.17 for a number 1.5 coverslip up to 0.20 to compensate for the "added glass" of the complex optical path results in increased image brightness and resolution of apical cell shapes (A-G). Further adjustment in this direction results in maintenance of image brightness but slightly decreased resolution of apical cell shapes (H-I). Correction collar adjusted in the opposite direction to 0.165 results in degraded image quality (J). (K) Enlarged regions of the same salivary gland acquired sequentially with correction collar settings of 0.17 and 0.20. Note the increased contrast between the salivary gland cells and the background and the increased resolution of apical cell shapes with the 0.20 setting.
DVDY21709SuppMovieS1.mpeg509KSupp Movie S1. High-resolution live imaging of wild-type salivary gland development. Full α-catenin-GFP and DsRed<SUP>N</SUP> signals are displayed to show the apical and basal dynamics of invagination and migration in the wild-type salivary gland. Contact with VM occurs at 30′ (corresponding to 0′ in Figure 2). Note that epithelial polarity and cell contacts are maintained throughout tube migration and reorientation.
DVDY21709SuppMovieS2.mpeg419KSupp Movie 2. Lumenal morphogenesis in the wild-type salivary gland. Only the brightest pixels of the α-catenin-GFP signal are displayed to highlight lumenal dynamics during wild-type salivary gland development. Contact with VM occurs at 30′ (corresponding to 0′ in Figure 2).
DVDY21709SuppMovieS3.mpeg433KSupp Movie 3. High-resolution live imaging of <I>ribbon</I> mutant salivary gland development. Full α-catenin-GFP and DsRed<SUP>N</SUP> signals are displayed to show the apical and basal dynamics of invagination and attempted migration in the <I>ribbon</I> mutant salivary gland. Contact with VM occurs at 30′ (corresponding to 0′ in Figure 2). Note that as in the wild-type salivary glands, epithelial polarity and cell contacts are maintained throughout tube migration and reorientation.
DVDY21709SuppMovieS4.mpeg379KSupp Movie 4. Lumenal morphogenesis in the <I>ribbon</I> mutant salivary gland. Only the brightest pixels of the α-catenin-GFP signal are displayed to highlight lumenal dynamics during <I>ribbon</I> mutant salivary gland development. Contact with VM occurs at 30′ (corresponding to 0′ in Figure 2).
DVDY21709SuppMovieS5.mpeg841KSupp Movie 5. High-resolution live imaging of wild-type tracheal dorsal trunk development. Full α-catenin-GFP and DsRed<SUP>N</SUP> signals are displayed to show the apical and basal dynamics of tube formation, elongation, and fusion in the wild-type tracheal dorsal trunk.
DVDY21709SuppMovieS6.mpeg512KSupp Movie 6. Lumenal morphogenesis in the wild-type tracheal dorsal trunk. Only the brightest pixels of the α-catenin-GFP signal are displayed to highlight lumenal dynamics during wild-type tracheal dorsal trunk development. Note that as in the salivary glands, epithelial polarity and cell contacts are maintained throughout tracheal tube migration and reorientation.
DVDY21709SuppMovieS7.mpeg980KSupp Movie 7. High-resolution live imaging of <I>ribbon</I> mutant dorsal trunk development. Full α-catenin-GFP and DsRed<SUP>N</SUP> signals are displayed to show the apical and basal dynamics of tube formation and attempted elongation during <I>ribbon</I> mutant dorsal trunk development.
DVDY21709SuppMovieS8.mpeg674KSupp Movie 8. Lumenal morphogenesis in the <I>ribbon</I> mutant dorsal trunk. Only the brightest pixels of the α-catenin-GFP signal are displayed to highlight lumenal dynamics during <I>ribbon</I> mutant dorsal trunk development. Note that as in the salivary glands and wild-type trachea, epithelial polarity and cell contacts are maintained throughout <I>ribbon</I> mutant tracheal tube migration and reorientation.

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