Frames from the middle boxed region in (A) showing three individual cells rearranging themselves from an oblique to a vertical stack (see also S4 Movie inset)
Frames from the middle boxed region in (A) showing three individual cells rearranging themselves from an oblique to a vertical stack (see also S4 Movie inset). nuclei, whose basal and apical widths were measured. = 43 cells, which were measured from four different biological specimens. D. Proportional basal cell abundance in bent epithelium versus flanking flat epithelium, calculated as (number cell type)/(total number of cells) in respective region. E. Average cell basal Griseofulvin and apical widths do Griseofulvin not significantly differ in bent Griseofulvin versus flat epithelium. F. Average cell base-to-apex width ratio shows a correlation of wedge-shaped cells with tissue invagination (note that the means of the ratios shown are not expected to be the same as the means of the averages shown in panel E). All error bars are +/- SDs. G. Phosphomyosin staining of an E12.5 tooth primordium (mesenchymal staining has been cropped out for clarity). H,I. Details of (G) showing that basal layer cells in the concave region have no apical phosphomyosin enrichment (H) or F-actin (I) enrichment.(TIF) pbio.1002405.s002.tif (1.9M) GUID:?FB38576B-BEC6-4D3A-B266-42EBA50068D6 S2 Fig: Arc tension recoil is produced specifically from elongated suprabasal cells, with no contribution from basal layer cells. A,B. Stills from timelapse video recording (S3 Movie) of a cut of similarly thickened, non-invaginating nasal epithelial tissue cut showing no recoil. CCF. Tooth placode frontal slices before and after a lateral cut in the presence of cytochalasin D (E,E) and blebbistatin (G, G) (no flexion). GCJ. Frontal tooth slice before and at the end of recoil produced by a suprabasal cell cut. I,J. Details of the boxed region in G and H, respectively. The dotted white lines mark the basal lamina, showing that, after indicated seconds, the basal lamina below the cut begins to kink (white arrowhead in J). K. Confocal Image of a frontal tooth slice fixed 2 min after performing a suprabasal cut experiment and stained with DAPI and AlexaFluor 488 Phalloidin. Only four of the most superficial suprabasal cell layers have been severed, leaving the basal palisade intact (thick white dashed line outlines the apical limit Griseofulvin of the basal palisade and thin dotted line outlines the basal lamina). Areas of this image were manipulated only to achieve a more uniform brightness. Size bars in G and I are 30 m, and Griseofulvin size bar in K is 100 m.(TIF) pbio.1002405.s003.tif (1.7M) GUID:?8A989F1C-0625-45C1-8E2A-7F7CD1B2833D S3 Fig: Shoulder nuclear shapes correlate with tissue curvature and tension. Shoulder cell nuclei were more apicobasally elongated than their non-shoulder neighbours and aligned with the direction of suprabasal cell intercalation because of their lean (arrows in ACE). Increased aspect ratio following shoulder lifting (curvature) starts on the buccal (cheek side) first at E12.5 (A,D) but then becomes symmetrical as the lingual shoulder lifts (D,E). This elongation is lost when the tissue-wide tension is released upon a lateral cut (dashed line in C), as indicated by loss of peaks in aspect ratio (F). Note that the latter relaxation is slower than that in suprabasal cells (2C4 h versus 10 min).(TIF) pbio.1002405.s004.tif (1.2M) GUID:?EC699451-5960-4964-A2FC-DDCD00D5250B S1 Movie: Explanted tooth epithelium bends autonomously (left), and manual re-extension and release of bent epithelium demonstrated tissue elasticity (right), related to Fig 1. The epithelium was microdissected from the mesenchyme using fine needles and cultured separately. Care was taken to explant the basal lamina intact. Timestamp is in mm:ss, 1 frame = 5 s.(AVI) pbio.1002405.s005.avi (19M) GUID:?18DA20CA-E9B7-4142-A23A-8500E79F9E5C S2 Movie: (Left) Lateral cut of a frontal tooth primordium slice showing rapid bending, related to Fig 1K and 1L, and (right) double cut of a frontal tooth primordium slice showing no flexing, related to Fig 1O and 1P. Timestamp is in mm:ss, 1 frame = 5 s.(AVI) pbio.1002405.s006.avi (1.1M) GUID:?DB7FE392-1E9F-4051-B5E1-B4242E75FA64 LPA receptor 1 antibody S3 Movie: Suprabasal cut of a frontal tooth primordium slice showing recoil, related to Fig 1M and 1N, and superficial cut of a frontal slice of thick nasal epithelium showing no recoil, related to S2A and S2B Fig (right). Timestamp is in mm:ss, 1 frame = 5 s.(AVI) pbio.1002405.s007.avi (314K) GUID:?CCC315E8-91D7-4976-B2BD-2419CFFC97C0 S4 Movie: Overview of 3 h confocal.