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. 2021 Sep 17;9:724310. doi: 10.3389/fcell.2021.724310

FIGURE 3.

FIGURE 3

Effect of dihydrotestosterone on outer root sheath cells regulated by dermal papilla cells in 2D and 3D co-culture models. (A) Effect of different concentrations of DHT on the proliferation of 2D- and 3D-cultured DPCs. (B) Relative ORS cell number in the four groups (control-2D, DHT-2D, control-3D, and DHT-3D). (C) Quantity of ORS cells cultured alone in the absence or presence of DHT (10 μM). Western blotting analysis of NCAM, β–catenin, and cleaved caspase-3 in the 2D- (D) and 3D-cultured (H) DPCs at 0 nM (control), 1 nM, 100 nM, and 10 μM DHT (D). Relative expression of NCAM, β–catenin (F), and cleaved caspase-3 in 2D (E–G) and 3D (I–K) DPCs. (L) Western blotting analysis of Ki67 and cleaved caspase-3 in 2D- and 3D-co-cultured ORS cells at 0 nM (control), 1 nM, 100 nM, and 10 μM DHT. Relative expression of Ki67 and cleaved caspase-3 in 2D (M,N) and 3D (O,P) ORS cells. The results were obtained using three replicates and presented as mean ± SD. ns, not significant; *p < 0.05; **p < 0.01; ***p < 0.001; and ****p < 0.0001 compared with the respective controls by one-way ANOVA and Student’s t-test. DHT, dihydrotestosterone; DPC, dermal papilla cell; NCAM, neural cell adhesion molecule; ORS, outer root sheath.