Journal: Nature Communications
Article Title: USP10 promotes glioma stem cell maintenance and glioblastoma growth by antagonizing DTX3L-mediated SATB2 ubiquitination
doi: 10.1038/s41467-025-67418-9
Figure Lengend Snippet: a IB analysis of SATB2, Myc, and USP10 expression in GSCs (T387) transduced with shNT, shUSP10-1, shUSP10-1+vector, or shUSP10-1 + SATB2-Myc. b IB analysis of SATB2, Myc, DTX3L, and Flag expression in GSCs (T387) transduced with vector, DTX3L-Flag, or DTX3L-Flag+SATB2-Myc. c Cell viability assay of GSCs (T387) transduced with shNT, shUSP10-1, shUSP10-1+vector, or shUSP10-1 + SATB2-Myc. n = 4 biological independent samples. d Cell viability assay of GSCs (T387) transduced with vector, DTX3L-Flag, or DTX3L-Flag+SATB2-Myc. n = 4 biological independent samples. e Quantification of the number and diameter of tumorspheres formed by GSCs (T387) expressing shNT, shUSP10-1, shUSP10-1+vector, or shUSP10-1 + SATB2-Myc. n = 4 biological independent cell cultures. f Quantification of the number and diameter of tumorspheres formed by GSCs (T387) expressing vector, DTX3L-Flag, or DTX3L-Flag+SATB2-Myc. n = 4 biological independent cell cultures. g In vitro limiting dilution analysis of the tumorsphere formations of GSCs (T387) expressing shNT, shUSP10-1, shUSP10-1+vector, or shUSP10-1 + SATB2-Myc. h In vitro limiting dilution analysis of the tumorsphere formations of GSCs (T387) expressing vector, DTX3L-Flag, or DTX3L-Flag+SATB2-Myc. i Kaplan-Meier survival curves of mice intracranially implanted with GSCs (T387) expressing shNT, shUSP10-1, shUSP10-1+vector, or shUSP10-1 + SATB2-Myc. n = 5–7 mice. j Kaplan-Meier survival curves of mice intracranially implanted with GSCs (T387) expressing vector, DTX3L-Flag, or DTX3L-Flag+SATB2-Myc. n = 6 mice. k Representative H&E images of mouse brains collected at day 28 after intracranially implantation with GSCs (T387) expressing shNT, shUSP10-1, shUSP10-1+vector, or shUSP10-1 + SATB2-Myc. Scale bar, 2 mm. l Representative H&E images of mouse brains collected at day 28 after intracranially implantation with GSCs (T387) expressing vector, DTX3L-Flag, or DTX3L-Flag+SATB2-Myc. Scale bar, 2 mm. Immunofluorescence images of Ki67 (green) and SOX2 (red) ( m ) with corresponding quantification of Ki67 + or SOX2 + cells ( n ) in tumor xenografts derived from GSCs (T387) expressing shNT, shUSP10-1, shUSP10-1+vector, or shUSP10-1 + SATB2-Myc. Scale bar, 50 μm. n = 5 tumors. Immunofluorescence images of Ki67 (green) and SOX2 (red) ( o ) with corresponding quantification of Ki67 + or SOX2 + cells ( p ) in xenografts derived from GSCs (T387) expressing vector, DTX3L-Flag, or DTX3L-Flag+SATB2-Myc. Scale bar, 50 μm. n = 5 tumors. Data are represented as mean ± s.d. One-way ANOVA analysis followed by Tukey’s test ( e , f , n , and p ), two-way ANOVA analysis followed by Tukey’s test ( c , d ), ELDA analysis (two-sided) for differences in stem cell frequencies was used in ( g , h ), log-rank test ( i , j ). Independent experiments were repeated three times, with similar results ( a , b , k , and l ).Source data are provided as a Source Data file.
Article Snippet: Lentiviral constructs expressing shRNAs against human USP10 , DTX3L , YY2 , and a non-targeting shRNA were cloned into pLKO.1-puro vector (Addgene, Cat# 8453).
Techniques: Expressing, Transduction, Plasmid Preparation, Viability Assay, In Vitro, Immunofluorescence, Derivative Assay