Supplementary Figure 4: Sebaceous gland function is maintained upon KrasG12D activation.

(a-c) Detection of Ki67 (green) and K14 (red) in WT tissue at indicated time-points. 3 animals per group. (d) Detection of FASN (green) and K14 (red) in KrasG12D expressing SG. 3 animals per group (e-h) Detection of BrdU (green) and K14(red) after a single pulse in WT and KrasG12D mutant mice at P7 and P56 n=3 animals per group. (i, j) Back skin tissue section from P23 stained with hematoxylin and Eosin from either WT or KrasG12D expressing mouse back skin at P23. Images representative of 3 animals per group. (k, l) Detection of lipids by Oil Red O (red) in back skin of WT and KrasG12D mutant mice at P23. Images representative of 3 animals per group. (m) SG size measurements according to Oil Red O surface area (µm²). Dots represent average measurements from independent animals (WT P7 n=3; P23 n=3; P56 n=4; P90 n=4; KrasG12D P7 n=3; P23 n=2; P56 n=3; P90 n=2). Data displayed are mean±S.E.M and statistical significance tested unpaired two-way students t-test. (n) Detection of clone (red) in a rendered confocal z-stack from KrasG12D tissue counterstained for IntegrinA6 (ItgA6, white). Image representative of 3 animals. (o) Fit of the basal clone size distributions with the same parameters as Fig. 5m, but incorporating a small refractory period until the next division, providing a slightly improved fit (solid lines). Basal clone size distribution of cohorts analysed at P7 (grey) and P23 (orange) are indicated (measurement are from Fig. 5i, j). (p) Basal (red) and suprabasal (blue) clone size distribution at P7 after a P2 induction, together with the corresponding best-fit predictions from a stochastic model of an equipotent imbalance progenitor population (used to extract the P2-P7 parameters of Fig. 5k). Lines and shaded areas represent best-fit predictions and 95% confidence interval respectively. (o,p) KrasG12D: P7-n=34 clones from 3 animals; P23-n=41 clones from 4 animals. (q) Numerical simulation of clone size distributions at P7 and P23 for a P2 induction based on a model of mixed KrasG12D and WT cells. Data is shown as dots (black: P7, red: P23) and models with different fractions f as lines (f=0% in green, f=20% in cyan, f=50% in purple). The model diverges from the data (in particular at P23) as f increases. Scale bars, 50 μm.