Fig. 2

Mutation of miR-200 sites in Zeb1 is sufficient to phenocopy miR-200 ablation. a Murine Zeb1 3′UTR, miR-200 site conservation, and mutated miR-200 site sequences. b Percent survival of RT2 mice with Zeb1 mutation (RT2, RT2_Zeb1^200H, RT2_Zeb1^200M, n = 42, 45, 26, respectively). c Random-fed blood glucose (RT2, RT2_Zeb1^200H, RT2_Zeb1^200M, n = 11, 11, 10, respectively). Dotted line represents onset of severe hypoglycemia. d Tumor burden in different age classes (box, 25th and 75th percentiles; central line, median; from left to right: n = 9, 4, 4, 4, 5, 11, 12 mice). e Representative insulin/glucagon and SV40 immunofluorescence staining of whole pancreas (left; insulin-negative tumors outlined; scale bar = 2 mm) and zoomed-in regions (right; all tumors outlined; scale bar = 100 μm). LN lymph node. f Percent tumors per grade (n = 5 mice per group; RT2, early-stage RT2_Zeb1^200M, late-stage RT2_Zeb1^200M, n = 55, 17, 16 tumors, respectively). g Percent insulin-positive and -negative tumors (n = 5 mice per group; RT2, early-stage RT2_Zeb1^200M, late-stage RT2_Zeb1^200M, n = 67, 12, 15 tumors, respectively). h Mean log₂FC (with 95% confidence intervals) of beta-cell-identity genes in RT2_Zeb1^200M vs. RT2 islets of 6-week-old mice. i Percent TUNEL-positive nuclei (n = 2 mice per group; RT2, early-stage RT2_Zeb1^200M, late-stage RT2_Zeb1^200M, n = 17, 21, 13 lesions, respectively). j Representative H&E and SV40 immunofluorescence staining of late-stage RT2_Zeb1^200M liver (scale bar = 2 mm). k Representative images of late-stage RT2_Zeb1^200H and RT2_Zeb1^200M mice. l Percent of mice with macrometastasis in difference age classes. c, i Data are plotted as mean ± SD. Significance was evaluated by b Mantel Cox test, c two-tailed t test with Holm−Sidak correction (vs. RT2), d, i one-way ANOVA with Dunnett’s multiple comparisons (vs. RT2) and h empirical Bayes method. *P ≤ 0.05; **P ≤ 0.01; ***P ≤ 0.001; ****P ≤ 0.0001