Figure 4.

Formation and growth of organoids from colon polyps of PKM2^ΔLgr5 mice. (a) Bright field images and quantification (number and size) of primary organoids from colon polyps of AOM/DSS-treated PKM2^ΔLgr5–Veh and –Tx mice. Surface area was measured at ≥50 organoids. Scale bar = 200 μm. (b) Bright field images and quantification (number and size) of secondary passaged organoids. Organoids were seeded at 5 × 10³ cells/well. Surface area was measured at ≥30 organoids. Scale bar = 500 μm. (c) Quantification by flow cytometry analysis of EpCAM⁺ Lgr5-GFP⁺ cells in colon polyps of AOM/DSS-treated PKM2^ΔLgr5–Veh and –Tx mice. (d) Bright field images and numbers of formed organoids from sorted single EpCAM⁺ Lgr5-GFP⁺ cells isolated from colon polyps of AOM-DSS treated PKM2^ΔLgr5-Veh and PKM2^ΔLgr5-Tx mice. Sorted cells were cultured in medium without R-spondin and Wnt3a. Scale bar = 200 μm. (e) Immunofluorescence staining of PKM1, PKM2, and Ki-67 in cancer organoids isolated from colon polyps of PKM2^ΔLgr5–Veh and –Tx mice. Scale bar = 100 μm. (f) Western blot analysis of cancer organoids from colon polyps of PKM2^ΔLgr5–Veh and –Tx mice. The full-length blots are presented in Fig. S6. The Data are representative of two independent experiments. All data are mean ± s.e.m. Statistical analyses were done by Student’s t-test. **p < 0.01.