In salivary gland epithelial cells, the actin cytoskeleton is organized primarily into cortical actin filaments at the cell perimeter (Figure 2C, 2D) in an E12 organ explant grown ex vivo for 0 or 24 hours. a tilt angle of less than 45 were eliminated as failed clefts.(TIF) pcbi.1003319.s003.tif (793K) GUID:?7A84EA83-9EBF-4E0E-8B81-5DD25CED807D Figure S4: Determination of relationship between focal point plasticity value, target distance D, and cleft-ECM contact energy (CM). (a) A simplified simulation was initialized with two 66 cells subjected to area and perimeter constraints, (b) A simulation was run for 1000MCS for varying Avarofloxacin values of D, CM and , and final cell distances were recorded. (c) Final stage of cell separation. (d) For each value of D selected, the and CM values required to achieve separation were saved and plotted. For these simulations, cell-cell contact energy value (CC) was kept constant at 10. A surface was fitted to these points in the form: This equation approximates the value required to achieve separation between two linked and opposing cleft cells under conditions in the single cleft simulation. It was used to select a range of focal point plasticity values that allowed us to examine the interplay between cleft-cell adhesion, cell-matrix adhesion, and mitosis rate.(TIF) pcbi.1003319.s004.tif (1.4M) GUID:?9E2B9E59-7D38-45F3-BA65-CACFA5FA2A6A Figure S5: Ratio of progressive to non-progressive clefts obtained during parametric search. (a) Mitotic rate (MR) variation from 0.5% to 5% (b) Focal point plasticity (FPP) variations from 0.5 to 30 (c) Cell-cell (CC) contact energy variation from 1 to 20 and (d) Cell-matrix (CM) contact energy variations from 1 to 5. For all the parameters, the corresponding ranges have been chosen based on the number of progressive clefts obtained in comparison to the number of non-progressive clefts and failed clefts.(TIF) pcbi.1003319.s005.tif (571K) GUID:?35953280-070E-4128-B712-23BEFB62C2DB Table S1: Cleft categorization during parametric search enabling choice of ranges of values for mitosis Avarofloxacin rate and focal point plasticity . Clefts were categorized as failed, progressive and non-progressive based on cleft depths measured from time-lapse videos of ex-vivo cultured explants.(DOCX) pcbi.1003319.s006.docx (16K) GUID:?38746707-B865-458E-899B-C74AEA167AE1 Table S2: Cleft categorization during parametric search enabling choice of ranges of values for cell-cell and cell matrix contact energies. Clefts were categorized as failed, progressive and nonprogressive based on cleft depths measured from time-lapse videos of ex-vivo cultured explants.(DOCX) pcbi.1003319.s007.docx (15K) GUID:?8A48D17F-DDBE-4CAD-85ED-3E2E6DF05479 Video S1: 20 hour time-lapse confocal movie of an E12 epithelial Avarofloxacin rudiment used for cleft depth measurements. (AVI) pcbi.1003319.s008.avi (8.3M) GUID:?B9875B76-2487-426B-AACE-B88D8B711172 Video S2: Example of a local cleft simulation with FPP?=?0 [T?=?10, Cell-cell adhesion at cleft cells?=?10, Cell-matrix adhesion at cleft cells?=?3, Mitosis rate?=?1%, Mitosis location?=?50% OCC, /50% IPC]. (AVI) pcbi.1003319.s009.avi (1.9M) GUID:?F7E28222-C24D-4BCF-8A41-A02701E8652C Video S3: Example of a GGH local salivary gland cleft simulation using base parameters: [T?=?10, Cell-cell adhesion at cleft cells?=?10, Cell-matrix adhesion at cleft cells?=?3, Mitosis rate?=?1%, Mitosis location?=?50% OCC/50% IPC, FPP (cleft cells)?=?10]. (AVI) pcbi.1003319.s010.avi (2.0M) GUID:?17AF1E75-386C-4B2F-AC9C-54668ECEE470 Video S4: Example of salivary gland organ level GGH simulation using base parameters [T?=?10, Cell-cell adhesion at cleft cells?=?10, Cell-matrix adhesion at cleft cells?=?3, Mitosis rate?=?1%, Mitosis location?=?50% OCC/50% IPC, FPP (cleft cells)?=?10]. (AVI) pcbi.1003319.s011.avi (8.0M) GUID:?9144760A-2F75-4DA9-B619-24262235BEE5 Video S5: Example of a local cleft simulation under conditions simulating ROCK I inhibition Avarofloxacin [T?=?10, Cell-cell adhesion at cleft cells?=?10, Cell-matrix adhesion at cleft cells?=?3, Mitosis rate?=?0.5%, Mitosis location?=?50% OCC/50% IPC, FPP (cleft cells)?=?1]. (AVI) pcbi.1003319.s012.avi (1.9M) GUID:?333BA538-0FA6-4112-9B44-ECDAB4DAECE5 Video S6: Example of a local cleft simulation under conditions simulating blebbistatin JV15-2 treatment [T?=?10, Cell-cell adhesion at cleft cells?=?10, Cell-matrix adhesion at cleft cells?=?3, Mitosis rate?=?1%, Mitosis location?=?50% OCC/50% IPC, FPP (cleft cells)?=?1]. (AVI) pcbi.1003319.s013.avi (2.0M) GUID:?BB8D1527-85AC-46AE-8275-FC60935E31BA Abstract Cleft formation during submandibular salivary gland branching morphogenesis is the critical step initiating the growth and development of the complex adult organ. Previous experimental studies indicated requirements for several epithelial cellular processes, such as proliferation, migration, cell-cell adhesion, cell-extracellular matrix (matrix) adhesion, and cellular contraction in cleft formation; however, the relative contribution of each of these processes is not fully understood since it is not possible to experimentally manipulate each factor independently. We present here a comprehensive analysis of several cellular parameters regulating cleft progression during branching.

In salivary gland epithelial cells, the actin cytoskeleton is organized primarily into cortical actin filaments at the cell perimeter (Figure 2C, 2D) in an E12 organ explant grown ex vivo for 0 or 24 hours