Figure 5. The block in sebocyte differentiation is lifted upon recovery of Notch activity.

(a) Representative hematoxylin and eosin (H&E) images of sebaceous glands (SGs) from mice (n=5 each) treated with aRW, aJ1, aJ2, and aJ1J2, 14 d post treatment. (b) Quantification of type of SG found after each treatment. The SGs were divided into three categories: normal SGs containing a characteristic number of sebocytes (normal SG), SGs containing a mix of sebocytes and basal-like cells (mixed SG), and SGs containing no sebocytes and only basal-like cells (no sebocyte SG). p-values: aJ1=0.535, aJ2=2.90E-87, aJ1J2=1.73E-92. Chi-square test used for statistical analysis. All treatments were compared against aRW. Total n of SGs quantified per treatment: aRW = 485, aJ1=430, aJ2=394, aJ1J2=364. Scale bars are 100 μm. (c,d) Representative co-stain images for N1ICD and fatty acid synthase (FASN) in SGs from mice (n=5 each) treated with aRW (c) and aJ2 (d), 7 d post-treatment. (e) Quantification of the percentage of N1ICD + cells in SGs from mice (n=5 each) treated with aRW and aJ2, 3 and 7 d post-treatment. Percentage was calculated by dividing the number of N1ICD+ cells by the total number of basal-like cells in each SG. p-values: for comparison between day 3 and day 7 for aRW treatment = 0.297, for comparison between day 3 and day 7 for aJ2 treatment = 1.00E-14. Total n of SGs quantified per treatment: aRW at 3 d = 13, aRW at 7 d = 13, aJ2 at 3 d = 15, aJ2 at 7 d=22. (f,g) Representative adipophilin staining in SGs from mice (n=5 each) treated with aRW (f) and aJ2 (g), 14 d post treatment. (h) Quantification of the number of cells expressing adipophiln in each SG, 7 and 14 d after treatment with aRW, and aJ2. p-values: for comparison between day 7 and day 14 for aRW treatment = 0.292, for comparison between day 7 and day 14 for aJ2 treatment = 1.38E-4. Total n of SGs quantified per treatment: aRW at 7 d = 18, aRW at 14 d = 19, aJ2 at 7 d = 22, aJ2 at 14 d=29. (i,j) Representative co-stain images for androgen receptor (AR) and FASN in SGs from mice (n=5 each) treated with aRW (i) and aJ2 (j), 14 d post-treatment. (k) Quantification of the number of cells expressing AR in each SG, 7 and 14 d after treatment with aRW and aJ2. p-values: for comparison between day 7 and day 14 for aRW treatment = 0.273, for comparison between day 7 and day 14 for aJ2 treatment=9.53E-09. Total n of SGs quantified per treatment: aRW at 7 d=12, aRW at 14 d=10, aJ2 at 7 d=18, aJ2 at 14 d=23. Student’s t-test used for statistical analysis. Error bars represent SEM. Scale bars are 25 μm.

Figure 5—figure supplement 1. The block in sebocyte differentiation is lifted upon recovery of Notch activity.

(Related to Figure 5). (a–c) Representative co-stain images for N1ICD and fatty acid synthase (FASN) in sebaceous glands (SGs) from mice (n=5 each) treated with aRW (a), aJ1 (b), and aJ1J2 (c), 7 d post-treatment. (d) Quantification of the percentage of N1ICD+ cells in SGs from mice (n=5 each) treated with aRW, aJ1, aJ2, and aJ1J2, 7 d post-treatment. Percentage was calculated by dividing the number of N1ICD+ cells by the total number of basal-like cells in each SG. p-values: aJ1=0.878, aJ2=3.42E-21, aJ1J2=5.60E-09. Total n of SGs quantified per treatment: aRW = 13, aJ1=13, aJ2=22, aJ1J2=13. (e–g) Representative adipophilin staining in SGs from mice (n=5 each) treated with aRW (e), aJ1 (f), and aJ1J2 (g), 14 d post-treatment. (h) Quantification of the number of cells expressing adipophilin in each SG, 14 d after treatment with aRW, aJ1, aJ2, and aJ1J2. p-values: aJ1=0.693, aJ2=3.40E-08, aJ1J2=0.003. Total n of SGs quantified per treatment: aRW = 19, aJ1=13, aJ2=29, aJ1J2=16. (i–k) Representative co-stain images for androgen receptor (AR) and FASN in SGs from mice (n=5 each) treated with aRW (i), aJ1 (j), and a1J2 (k), 14 d post-treatment. (l) Quantification of the number of cells expressing AR in each SG, 14 d after treatment with aRW, aJ1, aJ2. and aJ1J2. p-values: aJ1=0.248, aJ2=0.817, aJ1J2=0.032. Total n of SGs quantified per treatment: aRW=10, aJ1=7, aJ2=23, aJ1J2=18. Student’s t-test used for statistical analysis. Error bars represent SEM. Scale bars are 25 μm.