Fig 2: Expression of SOX2 in non-small cell lung carcinoma. (A and B) Expression of SOX2 in lung squamous carcinoma: (A) Negative (−), (B) strongly positive (+++). (C and D) Expression of SOX2 in lung adenocarcinoma: (C) Negative (−), (D) strongly positive (+++). (E) Expression of SOX2 in lung adenosquamous carcinoma: Weakly positive in adenocarcinoma cells (red arrow), strongly positive in squamous carcinoma (green arrow). (F) SOX2 shows a low expression in normal lung epithelial cells.

Fig 3: SATB2 is preferentially expressed by GSCs Immunofluorescence of SATB2 (green) and the GSC marker SOX2 or OLIG2 (red) on frozen tissue sections of human GBM surgical specimens. SATB2 is preferentially expressed by GSCs in human GBMs. Scale bar, 25 μm.Quantification of the fraction of SATB2+ cells in SOX2+ or OLIG2+ cells in human GBMs. More than 90% SOX2+ or OLIG2+ cells showed SATB2 staining. n = 3 GBMs.qPCR analysis of SATB2 mRNA expression in GSCs and matched non‐stem tumor cells (NSTCs) (n = 5).Immunoblot analysis of SATB2, SOX2, and OLIG2 expression in cell lysates of GSCs and matched NSTCs.Immunofluorescence of SATB2 (green) and SOX2 (red) in T3359 GSCs and matched NSTCs. Scale bar, 50 μm.qPCR analysis of SATB2 mRNA expression in GSCs and neural progenitor cells (NPCs) (n = 3).Immunoblot analysis of SATB2 and SOX2 expression in cell lysates of GSCs and NPCs.Immunoblot analysis of SATB2, GSC marker SOX2, and differentiation marker GFAP expression during serum‐induced GSC differentiation. Data information: Data are represented as mean ± SD. **P < 0.01, ***P < 0.001, Mann–Whitney test (C) or one‐way ANOVA analysis followed by Tukey’s test (F). Exact P values are reported in Appendix Table S3.

Fig 4: Association between SOX2 expression and prognosis of non-small cell lung carcinoma. Survival analysis revealed that the prognosis of patients of the high expression group was better (P=0.008) and the 5-year survival rate was higher (high vs. low: 55.5 vs. 35.3%).

Fig 5: Ectopic expression of FOXM1 rescued the decreased GSC proliferation and GBM growth caused by SATB2 disruption AImmunoblot analysis of FOXM1 and SATB2 expression in T3359 GSCs transduced with FOXM1 or vector control in combination with shNT or shSATB2.BCell viability assay of T3359 GSCs transduced with FOXM1 or vector control in combination with shNT or shSATB2 (n = 5). Ectopic expression of FOXM1 restored the cell proliferation impaired by SATB2 silencing.CTumorsphere number of T3359 GSCs transduced with FOXM1 or vector control in combination with shNT or shSATB2 (n = 5). Ectopic expression of FOXM1 restored the tumorsphere formation of GSCs impaired by SATB2 silencing.D, E In vivo bioluminescent images (D) or quantification (E) of the tumor xenografts derived from luciferase‐labeled T3359 GSCs transduced with FOXM1 or vector control in combination with shNT or shSATB2 (shNT: n = 5 mice; shNT + FOXM1: n = 4 mice; shSATB2: n = 5 mice; shSATB2 + FOXM1: n = 5 mice). Representative images on day 21 posttransplantation are shown. Ectopic expression of FOXM1 in GSCs expressing shSATB2 markedly restored GBM tumor growth.FKaplan–Meier survival curves of mice intracranially implanted with T3359 GSCs transduced with FOXM1 or vector control in combination with shNT or shSATB2 (shNT: n = 8 mice; shNT + FOXM1: n = 7 mice; shSATB2: n = 6 mice; shSATB2 + FOXM1: n = 8 mice). Median survival: shNT, 30 days; shNT + FOXM1, 28 days; shSATB2, 50.5 days; shSATB2 + FOXM1, 34 days. Ectopic expression of FOXM1 in GSCs expressing shSATB2 markedly attenuated the increased survival of mice bearing the GSC‐derived GBMs.GImmunofluorescence of Ki67 (Green) in xenografts derived from T3359 GSCs transduced with FOXM1 or vector control in combination with shNT or shSATB2 (shNT: n = 6 tumors; shNT + FOXM1: n = 5 tumors; shSATB2: n = 6 tumors; shSATB2 + FOXM1: n = 6 tumors). Scale bar: 40 μm.HQuantification of Ki67 positive cells in xenografts derived from T3359 GSCs transduced with FOXM1 or vector control in combination with shNT or shSATB2 (shNT: n = 6 tumors; shNT + FOXM1: n = 5 tumors; shSATB2: n = 6 tumors; shSATB2 + FOXM1: n = 6 tumors).IImmunofluorescence of SOX2 (Red) in xenografts derived from T3359 GSCs transduced with FOXM1 or vector control in combination with shNT or shSATB2 (shNT: n = 6 tumors; shNT + FOXM1: n = 5 tumors; shSATB2: n = 6 tumors; shSATB2 + FOXM1: n = 6 tumors). Scale bar: 40 μm.JQuantification of SOX2 positive cells in xenografts derived from T3359 GSCs transduced with FOXM1 or vector control in combination with shNT or shSATB2 (shNT: n = 6 tumors; shNT + FOXM1: n = 5 tumors; shSATB2: n = 6 tumors; shSATB2 + FOXM1: n = 6 tumors). Data information: Data are shown as mean ± SD. **P < 0.01, ***P < 0.001, ****P < 0.0001, two‐way ANOVA analysis followed by Tukey’s test (B), one‐way ANOVA analysis followed by Tukey’s test (C, E, H, and J) or log‐rank test (F). Exact P values are reported in Appendix Table S3.