Fig 2: Loss of histone H3 lysine 27 trimethylation accompanies SCC transition.(a) Western blotting analysis performed on whole-cell extracts from tumours of the indicated genotypes and histologies. Histone H3 lysine 27 tri-methylation (H3K27me3) is markedly lower in SCC lesions; total histone H3 is the loading control. (b) Western blotting analysis performed on whole-cell extracts from tumours of the indicated genotypes and histologies. p63 and LKB1 confirm the histologies and genotypes of the lysates, and while EZH2 in more highly expressed in SCC tumours, the essential PRC2 subunit EED is absent in the SCC lesions. ß-Actin is the loading control. (c) Immunohistochemisty for H3K27me3 on a panel of mouse lung tumours of the indicated histologies from KRAS/Lkb1, Pten/Lkb1/p53, and Pten/Lkb1 mice, scale bar, 50 µm. (d) Quantification of nuclear staining by dot analysis with Nikon software; data are mean±s.e.m. measured on serially stained sections, n=6–10. (e) Immunohistochemisty for H3K27me3 on a panel of human mixed adenocarcionomas, pure ADC and pure SCCs. Scale bar, 50 µm. (f) Quantification of nuclear staining, for H3K27me3 n=6 ADSCC, 14 ADC, 9 SCC, for EZH2 n=6 ADSCC, 7 ADC, 5 SCC, data are mean±s.e.m. measured on serially stained sections. P values represent 2 tailed t-test. See also Supplementary Fig. 3a–c.

Fig 3: Activation of YAP signaling by XMU-MP-1 promotes OE regeneration and functional recovery of the olfactory system after injury(A) Immunohistochemistry detected the expression of YAP in the OE of control- and XMU-MP-1-treated 2-month-old C57BL/6 mice at 0, 3, and 7 dpl.(B) Quantitative analysis of the relative YAP density as shown in (A) (n = 6 sections from 6 mice per group).(C) Western blot detected the expression of p-MOB1, MOB1, p-YAP, and YAP in OE of control- and XMU-MP-1-treated 2-month-old C57BL/6 mice at 7 dpl.(D and F) Quantitative analysis of the relative p-MOB1/MOB1 (D), p-YAP/YAP (E), and YAP (F) levels as shown in (C) (normalized with control group, n = 8 blots from 4 mice per group, Student’s t test).(G) H&E staining of the OE obtained from control- and XMU-MP-1-treated 2-month-old C57BL/6 mice at 0, 3, and 7 dpl.(H) Quantitative analysis of the thickness of OE as shown in (G) (n = 12 sections from 6 mice per group).(I) Quantitative analysis of the time to locate food of control-treated (n = 10 mice) and XMU-MP-1-treated 2-month-old C57BL/6 mice (n = 13 mice) in the buried pellet test at 7 dpl (Student’s t test).(J and K) Double immunostaining of Ki67 (green) and p63 (red) (J) or PH3 (green) and K5 (red) (K) in the OE of control- and XMU-MP-1-treated 2-month-old C57BL/6 mice at 0 and 3 dpl.(L and M) Quantitative analysis of the percentage of both Ki67+ and p63+ (L) or PH3+ and K5+ (M) cells over p63+ or K5+ HBCs as shown in (J) and (K) (n = 13 sections from 6 mice per group).(N) Immunostaining of GAP43 (green) in the OE of control- and XMU-MP-1-treated 2-month-old C57BL/6 mice at 7 dpl.(O) Quantitative analysis of the density of GAP43+ cells as shown in (N) (n = 12 sections from 6 mice per group, Student’s t test).Images of selected regions are shown at higher magnification. Data are mean ± SEM, two-way ANOVA with Bonferroni’s post tests unless otherwise indicated, compared with control group; *p < 0.05, **p < 0.01, ***p < 0.001. Scale bars, 20 µm.

Fig 4: Inhibition of S1PR2 impairs OE regeneration and functional recovery of the olfactory system after injury(A) H&E staining of the OE obtained from control- and JTE013-treated 2-month-old C57BL/6 mice at 0, 5, and 14 dpl.(B) Quantitative analysis of the thickness of OE as shown in (A) (n = 7 sections from 4 mice per group).(C) Percentage of mice that found cookies over total observed control-treated (n = 8 mice) or JTE013-treated 2-month-old C57BL/6 mice (n = 7 mice) in the buried pellet test at 14 dpl (Student’s t test).(D) Quantitative analysis of the time to locate food in control-treated (n = 8 mice) or JTE013-treated 2-month-old C57BL/6 mice (n = 7 mice) at 14 dpl in the buried pellet test (Student’s t test).(E and F) Double immunostaining of Ki67 (green) and p63 (red) (E) or PH3 (green) and K5 (red) (F) in the OE of control- and JTE013-treated 2-month-old C57BL/6 mice at 0, 3, and 5 dpl.(G and H) Quantitative analysis of the percentage of both Ki67+ and p63+ cells (G) or PH3+ and K5+ (H) cells over p63+ or K5+ HBCs as shown in (E) and (F) (n = 10 sections from 4 mice per group).(I) Double immunostaining of GAP43 (green) and OMP (red) in the OE of control- and JTE013-treated 2-month-old C57BL/6 mice at 0, 7, and 14 dpl.(J and K) Quantitative analysis of the density of GAP43+ (J) or OMP+ (K) cells at 0, 3, 7, and 14 dpl (n = 6 sections from 4 mice per group).Images of selected regions are shown at higher magnification. Data are mean ± SEM, two-way ANOVA with Bonferroni’s post tests unless otherwise indicated, compared with control group; *p < 0.05, **p < 0.01, ***p < 0.001. Scale bars, 20 µm.

Fig 5: Immunocytochemical analysis of the CTNNB1 localization by RBM24. Immunocytochemical analysis was performed on (A) HepG2 and (B) Hep3B cells treated with 25 mM LiCl and/or RBM24-pcDNA or control pcDNA vectors, and cells were collected to assess the induction of CTNNB1 nuclear translocation using DAPI (nucleus), CTNNB1 and TP63 staining and merging them. Change in TP63 expression was indicated according to CTNNB1 nuclear translocation and upon treatment with siCTNNB1. Scale bar, 20 µm. CTNNB1, ß-catenin; RBM24, RNA-binding protein 24; TP63, tumor protein 63; si-, small interfering.