Figure 3: Hmga1 deficiency disrupts ISC function in 3D organoids.

(a) Representative images of organoid formation after silencing Hmga1 by delivery of lentiviral vector expressing shRNA targeting Hmga1 (knockdown or KD) or control lentivirus (Cont) are shown. Scale bars, 50 μm. (b) Relative Hmga1 expression (mean±s.d.) was assessed from two experiments performed in triplicate (via qPCR) in organoids transduced with lentivirus (Cont versus KD) at day 10. Gapdh was used to control for loading. **P<0.01; two-tailed Student’s t-test. (c) PSA (mean±s.d.) was assessed in organoids transduced with lentivirus (Cont versus KD) at the indicated time points (n=20 organoids per group). *P<0.05, **P<0.01; two-tailed Student’s t-test. (d) Representative image of organoid formation±knockdown of Hmga1 via an inducible lentiviral vector expressing red fluorescent protein (RFP) and shRNA targeting Hmga1 are shown. Doxcyline (0.5 μg ml⁻¹) was used to induce RFP and shRNA. Scale bars, 50 μm. (e) Relative Hmga1 expression (mean±s.d.) in organoids with or without induction of lentivirus shRNA targeting Hmga1 was assessed from two experiments performed in triplicate (via qPCR); Gapdh was used to control for loading. *P<0.05, **P<0.01; two-tailed Student’s t-test. (f) PSA (mean±s.d.) of organoids with or without induction of Hmga1 knockdown at the indicated time points are shown (n=20 organoids per group from two different virus transduction experiments). **P<0.01; Mann–Whitney test.