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. 2023 Jun 25;114(9):3636–3648. doi: 10.1111/cas.15882

FIGURE 2.

FIGURE 2

Inhibition of the bone morphogenetic protein (BMP) pathway suppresses colorectal cancer (CRC) organoid growth. (A) Dose–response curves of LDN193189 (LDN) in 18 CRC organoid lines cultured in growth factor‐free medium. (B) Bright‐field images of CRC organoids cultured in the presence (lower panels) or absence (Ctrl, upper panels) of 0.1 μM LDN for 5 days. Scale bar, 100 μm. (C) Cell viability assay results for CRC organoids. Organoids were treated with 0.1 μM LDN or 100 ng/mL Noggin. Number of replicates is indicated below the organoid names. Data are presented as mean + SD. Statistical comparisons were made for each control condition. (D) Immunoblotting analysis of CRC organoids cultured in the presence (LDN) or absence (−) of 0.1 μM LDN for the indicated time (h). Proteins were detected with indicated Ab. (E–G) Cell viability assay results for C45 organoids. Organoids were treated with 0.1 μM LDN with or without 50 ng/mL epidermal growth factor (EGF) (n = 5 for each condition, E) or heregulin (HRG) at the indicated concentration (n = 3 for each condition, F). Data are presented as mean + SD. (G) Dose–response curves of LDN in 16 CRC organoid lines cultured in StemPro hESC medium. *p < 0.01, t‐test with Bonferroni correction. ACTB, β‐actin; EGFR, EGF receptor; NS, not significant.