Supplementary Figure 4: Quantitative analysis of incisor epithelial kinetics using the TetOn-H2B-GFP system. | Nature Cell Biology

Supplementary Figure 4: Quantitative analysis of incisor epithelial kinetics using the TetOn-H2B-GFP system.

From: A large pool of actively cycling progenitors orchestrates self-renewal and injury repair of an ectodermal appendage

Supplementary Figure 4

a, Frontal images and µCT rendering of the hemi-mandible from control (top), tetOff (middle) and tetON (bottom)-H2B-GFP mice. tetOff (middle) mice possessed chalky white incisors with blunted tips and missing enamel (yellow arrowhead), as compared to the yellow, sharp and intact incisors in controls (top) and TetOn-H2B-GFP (bottom) mice. µCT analysis shows that enamel is compromised (pink arrowheads) in the tetOff-H2B-GFP mice. Coronal sections at a region distal to the first molar [red dotted lines in (b)] show reduced density (blue arrowheads) and defective enamel (red arrowhead) in TetOff-H2B-GFP mice. Scale bars, 5 mm (in a- 3D),1 mm (in coronal section). b, Quantification of the incisor volume (top) and enamel density (bottom) assessed by µCT. N=3 animals. Bar charts indicate the mean of samples and error bars represent ± S.D. of the mean. Data were analyzed using unpaired t-test with Welch's correction. Significance was taken as P<0.05 with a confidence interval of 95%. * P< 0.05. c, Immunostaining for GFP in the incisor growth region. Prior to Dox induction, the incisor epithelium in TetOn-H2B-GFP is GFP-negative. Scale bar, 100 µm. d, Immunostaining for GFP in the incisor growth region of TetOn-H2B-GFP mice pulsed (Dox) between postnatal day (P) 2 and 4 and chased for 8 weeks. The OEE and the adjacent SR maintained high levels of GFP, which persisted for at least 8 weeks. Scale bars, 100 µm. e, Immunostaining for PAX7 (red) and GFP (green) in the anterior tibialis muscle of TetOn-H2B-GFP mice. Scale bars, 10 µm. f, Immunostaining for GFP in the incisor growth region of TetOn-H2B-GFP mice pulsed (Dox) for 21 days starting at 8 weeks of age and chased for 6 (top) and 12 (bottom) months. The IEE and the rest of the proliferating regions of the incisor epithelium are negative for GFP (yellow insets), while the lower regions of the OEE and the adjacent SR (orange insets) maintain high levels of GFP. The layers of cells ventral to the SI and the distal ameloblasts layer (pink insets) also contain GFP-positive cells. Scale bars, 100 µm.

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