Fig. 3. SCM-induced cystic organoid phenotype is dependent on Ptk7.

a Western blot for sPtk7 in conditioned media collected from quiescent and senescent MEFs treated with vehicle or inhibitors for 24 h. Conditioned media corresponding to the same number of cells was loaded. b Representative images of organoids cultured in corresponding conditioned media. Scale bar, 200 μm. c Quantification of data from (b). The percentage of cystic organoids is shown. d Western blot for sPtk7 in conditioned media collected from senescent Ptk7^+/fl and Ptk7^fl/fl fibroblasts. Conditioned media corresponding to the same number of cells was loaded. e Representative images of organoids cultured in corresponding conditioned media. Scale bar, 200 μm. f Quantification of data from (e). g Representative images of organoids cultured in conditioned media together with IgG (5 μg/ml) or increasing concentration of anti-Ptk7 antibodies (1, 2, and 5 μg/ml). Scale bar, 200 μm. h Quantification of data from (g). i Western blots of full-length and secreted Ptk7 and MMP14 in whole-cell extract of quiescent and senescent primary intestinal fibroblasts or conditioned media collected from the fibroblasts. Tubulin was used as loading control. j Representative images of organoids co-cultured with quiescent or senescent intestinal fibroblasts in the presence of IgG or anti-Ptk7. Scale bar, 200 μm. k Quantification of data from (j). In (c), (f), (h), and (k), data are presented as means ± s.d. One-way ANOVA followed by Tukey’s post-hoc multiple comparison tests. Each dot represents a result from organoids established from a mouse, n = 3 in (c) and (h), n = 4 in (f), and n = 5 in (k). See also Fig. S2. Source data are provided as a Source Data file.