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

FIGURE 3.

FIGURE 3

Bone morphogenetic protein (BMP) inhibition induces LRIG1‐mediated degradation of epidermal growth factor receptor (EGFR). (A,B) Gene expression of EGFR (A) and LRIG1 (B) in colorectal cancer organoids treated with LDN193189 (LDN) or Noggin. Expression level was normalized to control (Ctrl) in each organoid. n = 3 for each condition. Data are presented as mean + SD. Statistical comparisons were made for each control condition. (C) Co‐immunoprecipitation (IP) analysis of lysates from C45 organoids treated with 0.1 μM LDN compared to untreated control (Ctrl). Samples were immunoblotted (IB) with EGFR, ubiquitin, and β‐actin (ACTB) Abs. Input lysates were used as loading controls. (D) Immunoblotting analysis of C45 organoid cultured for 48 h with LDN at the indicated concentration (nM) in the presence (+) or absence (−) of 5 nM bafilomycin A1. Proteins were detected with indicated Abs. (E) Cell viability assay results for C45 organoids. Organoids were treated with 0.1 μM LDN with or without 5 nM bafilomycin A1 (BAF). Data are presented as mean + SD. N = 3. (F) Immunoblotting analysis of C45 organoid cultured with 0.1 μM LDN or 250 ng/mL Noggin for 48 h. Proteins were detected with indicated Abs. *p < 0.05, **p < 0.001, t‐test with Bonferroni correction. HER, human epidermal growth factor receptor; NS, not significant.