Extended Data Fig. 2: Basal progenitors are increased after androgen treatment.

a) Quantification of TBR2+ cells per mm² progenitor zone (see Extended Data Fig. 1a) at 35d. b) Image of the ventricular zone/subventricular zone (VZ/SVZ) border in XX 35d organoids stained for Ki67 (green, white) and TBR2 (white). Cells positive for both Ki67 and TBR2 are indicated with magenta arrowheads. Double positive mitotic cells are indicated with yellow arrowheads. A white dashed line delineates VZ/SVZ border with the ventricular zone below. c) Immunostaining for proliferation marker Ki67 (white) on XX 35d control (left) and DHT treated (right) organoids. Dashed yellow lines represent the apical surface (bottom line) and the VZ/SVZ boundary. Note the difference in Ki67+ cells in the SVZ of DHT-treated organoids. d) Quantification of Ki67/TBR2 double positive cells, out of all TBR2+ cells, in XX 35d and 52d organoids. e) Immunostaining for HOPX (fire LUT) of XX 52d organoids. Yellow dashed line indicates the ventricular surface. Note the HOPX signal in radial glia. Images are single, 1.2 µm optical planes. f) Quantification of HOPX+ basal radial glia per mm² SVZ of XX 52d organoids. g) Immunostaining for phosphorylated histone H3 (PH3) (green) on XX 35d and XY 52d old organoids. Co-stained with DAPI (white). h) Quantification of apical mitotic cells (PH3+), normalized per mm length of ventricle in male (XY) and female (XX) organoids, at 35d and 52d, treated with DHT, T and E. i) Quantification of ventricular length in male (XY) and female (XX) organoids, at 35d and 52d, treated with DHT, T and E. Scale bars: g) 100 µm, e) 50µm, b), c) 20µm. See Methods for details of statistics and Supplementary Table 5 for details of n numbers.