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SC_11-0696_sm_supplFigure1.tiff1675KFigure S1. BM18 xenografts contain a human epithelial compartment and a mouse stromal compartment. (A and B): Fluorescence images of Hoechst-33528 stained sections (left panels) and corresponding H&E stainings on consecutive sections (right panels) of BM18 xenografts in (A) intact hosts and (B) in castrated hosts 21 days after surgery. Dashed lines delineate stromal cells with mouse specific punctate nuclear staining from epithelial cells with human specific diffuse nuclear staining. Scale bar is 50μm.
SC_11-0696_sm_supplFigure2.pdf131KFigure S2. Neuroendocrine cells in BM18 tumors are CK18-positive and AR-negative. Histological sections of BM18 xenografts in (A) intact host and in (B-C) castrated host 23 days after surgery were derived either from (A-B) frozen BM18 tumors or from (C) paraffin-embedded BM18 tumors. (A-B): Immunofluorescent co-localization of CK18 (left panels) and SYP (middle panels) co-staining. Right panels: merged pictures of CK18 (green), SYP (red) and DAPI (blue) staining. Arrowheads in A indicate SYP-positive cells. (C): Immunohistochemical co-localization of AR and CHGA on consecutive sections derived from BM18 xenografts castrated host 23 days after surgery. Arrowheads indicate AR-negative/CHGA-positive cell. Insets represent a higher magnification of selected areas. Scale bar is 50μm.
SC_11-0696_sm_supplFigure3.tiff2439KFigure S3. ALDH1-positive cells in BM18 xenografts in castrated hosts are ALDH1A1-positive. (A): mRNA expression levels of different ALDH genes in BM18 xenografts in intact and castrated hosts 14 days after surgery. Values are shown as mean ± SD and are relative to β-actin (ACTB) mRNA levels. *,**: Significantly different (*: p<0.05, **: p<0.01). (B-E): Immunohistochemical co-localization of (B, D) ALDH1 and (C, E) ALDH1A1 on consecutive sections derived from BM18 xenografts in (B-C) intact and castrated host 23 days after surgery (D-E). Arrowheads indicate ALDH1A1- positive cells. Insets represent a higher magnification of selected areas. Scale bar is 50μm.
SC_11-0696_sm_supplFigure4.pdf140KFigure S4. ALDH1-positive cells and NANOG-positive cells have a distinct expression of luminal markers. (A and B): Immunohistochemical co-localization of luminal markers (AR, PSA, NKX3.1) and (A) ALDH1 or (B) NANOG on consecutive sections derived from BM18 xenografts in castrated hosts. Filled arrowheads indicate ALDH1pos cells in A and NANOGpos cells in B. Open arrowheads indicate PSApos cells in the middle panel in A and NKX3.1high cells in lower panels in A and B. Scale bar is 50μm.
SC_11-0696_sm_supplFigure5.tiff1692KFigure S5. ALDH1-positive cells in BM18 xenografts in castrated hosts do not co-localize with NANOG-positive cells. (A-C): Immunohistochemical co-localization of ALDH1, NANOG and CD44 on consecutive sections derived from BM18 xenografts in castrated hosts. Filled arrowheads indicate ALDH1pos cells and open arrowheads indicate NANOGpos cells. Scale bar is 50μm.
SC_11-0696_sm_supplFigure6.tiff2339KFigure S6. Castration-resistant BM18 cancer cells re-express luminal differentiation marker genes after androgen-replacement. mRNA expression in BM18 xenografts in castrated hosts 72 hours after androgenreplacement. Values are derived from three to four different xenografts and are shown as relative to xenografts in castrated hosts on a logarithmic scale. *,**: Significantly different from castrated host (*: p<0.05, **: p<0.01).
SC_11-0696_sm_supplFigure7.pdf132KFigure S7. Two distinct cell populations contribute differently to BM18 tumor growth in either intact or androgen-replaced, castrated hosts. (A): percentage of ALDH1-postive and –negative (ALDHpos, ALDHneg) Ki67- positive (Ki67pos) cancer cells in BM18 xenografts in intact or castrated hosts at indicated times after androgen-replacement. (B): Immunofluorescent colocalization of Ki67 (red) and ALDH1 (green) in a section of a BM18 xenograft in intact host. Blue: DAPI nuclear counterstaining. Panels I – XV represent different fields. Arrowhead in panel XI indicates the only weakly Ki67- positive/ALDH1-positive cell found in an entire xenograft section. Scale bar is 50μm.
SC_11-0696_sm_supplFigure7.tiff4191KFigure S7. Two distinct cell populations contribute differently to BM18 tumor growth in either intact or androgen-replaced, castrated hosts. (A): percentage of ALDH1-postive and –negative (ALDHpos, ALDHneg) Ki67- positive (Ki67pos) cancer cells in BM18 xenografts in intact or castrated hosts at indicated times after androgen-replacement. (B): Immunofluorescent colocalization of Ki67 (red) and ALDH1 (green) in a section of a BM18 xenograft in intact host. Blue: DAPI nuclear counterstaining. Panels I – XV represent different fields. Arrowhead in panel XI indicates the only weakly Ki67- positive/ALDH1-positive cell found in an entire xenograft section. Scale bar is 50μm.
SC_11-0696_sm_supplTable1.pdf55KSupplementary Table 1
SC_11-0696_sm_supplTable2.pdf56KSupplementary Table 2

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