Journal: bioRxiv

Article Title: DTX3L–PARP9 co-evolve as a single adaptive unit linking ubiquitination and ADP-ribosylation in antiviral immunity

doi: 10.64898/2026.04.23.720386

Figure Lengend Snippet: a) Domain architecture of Deltex family proteins, PARP9, and PARP14. Canonical domains are mapped along each protein using domain boundaries defined in InterPro, showing common and differing features between proteins from the same families. b) Phylogenetic representation of gene presence and absence across 21 metazoan classes. Filled cells indicate the presence of an ortholog, while empty cells denote absence. Deltex family proteins are shown in red (lighter red for inferred ancestral Deltex proteins) and PARP proteins in blue. Numbers on the right represent the number of species with the represented profile of presence and absence over the total number of species analyzed in the class. c) Maximum-likelihood phylogeny of the Deltex family reconstructed from full-length protein sequences. Major clades corresponding to DTX1–4 and DTX3L are labeled, with nodes support (bootstrap values) indicated by circles when > 0.7. d) Predicted domain architectures of DTX3L orthologs across representative metazoan species, illustrating conserved features and lineage-specific differences, and shared domains with PARP14. e) Graphical representation of the synteny of the canonical DTX3L locus across representative metazoan species and, the alternate PARP9-associated locus, showing the emergence of a conserved DTX3L-PARP9 locus following the diversification of dipnoi (lungfishes). f) Correlation between transcriptional expression (z) of DTX3L and PARP9 and their intergenic distance (Log10 genomic distance +1). Analysis was performed for different species ( Rattus norvegicus, Mus musculus, Homo sapiens, Macaca mulatta, Gallus gallus, Canis lupus and Felis catus ) show distinct intergenic distance, using available co-expression data from COXPRESdb. Significance of the slope was tested with Spearman test (ρ = −0.8, p-value = 0.029).

Article Snippet: This pattern is noteworthy given that, within the DELTEX family, DTX3L and DTX3 are the only members harboring ssNA-ubiquitinating activity ( ).

Techniques: Labeling, Expressing

Journal: bioRxiv

Article Title: DTX3L–PARP9 co-evolve as a single adaptive unit linking ubiquitination and ADP-ribosylation in antiviral immunity

doi: 10.64898/2026.04.23.720386

Figure Lengend Snippet: a) Maximum likelihood phylogenetic trees of mammalian (primates, rodents, artiodactyls, bats and carnivores) DTX3L, showing branches under significant positive selection (p-value <0.05, in red). b) Summary of BUSTED (Branch-site Unrestricted Statistical Test for Episodic Diversification) and models M0, M1, M2, M7, and M8 implemented in Bio++ and codeml (PAML) with nested model comparisons (M1 vs. M2 and M7 vs. M8) assessed by likelihood ratio tests (LRTs) on DTX3L across mammalian orders. Colored boxes indicate models detecting a significant signal of positive selection and barplots indicate the proportion of all positively selected sites (Unique PSS) and Significant PSS (shared by at least two models). c) Significant positively selected sites mapped on the linear representation of DTX3L protein structure for each mammalian order. Red bars indicated the PSS shared by at least three mammalian orders. The dotted line box show the stretch of shared PSS. Codon numbering is based on the reference sequences for each order: artiodactyla - Bos mutus , carnivora - Canis lupus , chiroptera - Pteropus giganteus , primata - Homo sapiens , rodentia - Mus musculus ) . d) Alignments of positively selected sites shared by at least three mammalian orders, highlighting amino acid changes in mammalian DTX3L. The cladogram on the left shows the phylogenetic relationships among mammalian species. e) Predicted three-dimensional structures of DTX3L generated using AlphaFold2 for three representative mammalian species : H. sapiens, B. mutus , and P. giganteus . f) Close-up view of the DTC domain of DTX3L, where shared significant PSS across at least three orders are shown in red.

Article Snippet: This pattern is noteworthy given that, within the DELTEX family, DTX3L and DTX3 are the only members harboring ssNA-ubiquitinating activity ( ).

Techniques: Selection, Generated

Journal: bioRxiv

Article Title: DTX3L–PARP9 co-evolve as a single adaptive unit linking ubiquitination and ADP-ribosylation in antiviral immunity

doi: 10.64898/2026.04.23.720386

Figure Lengend Snippet: a) Summary of BUSTED and models M0, M1, M2, M7, and M8 implemented in Bio++ and Codeml (PAML) with nested model comparisons (M1 vs. M2 and M7 vs. M8) assessed by likelihood ratio tests (LRTs) on PARP9 and PARP14. Colored boxes indicate models detecting a significant signal of positive selection and barplots indicate the proportion of all positively selected sites (Unique PSS) and Significant PSS (shared by at least two models). b) Significant positively selected sites mapped on the linear representation of PARP9 and PARP14 protein structures for each mammalian orders. c) Alignments of positively selected sites shared by at least three mammalian orders, highlighting amino acid changes in mammalian PARP9. The cladogram on the left shows the phylogenetic relationships among mammalian species. Codons with asterisk indicate the most rapidly evolving codons (BEB > 0.99 and detected by at least four methods). d) Predicted 3D docking structures of DTX3L-PARP9 complex from human using Colabfold (pLDDT>70), showing the spatial conformation of DTX3L RING-DTC and PARP9 macrodomain (left) and the enrichment of positively selected sites in both domains (right). e) Predicted 3D docking structures of DTX3L-PARP9 complex from human, with a close view of the positively selected sites in the interface (green). f-g ) Similar analyses for the bat species P. giganteus as described for c-d.

Article Snippet: This pattern is noteworthy given that, within the DELTEX family, DTX3L and DTX3 are the only members harboring ssNA-ubiquitinating activity ( ).

Techniques: Selection

Journal: bioRxiv

Article Title: DTX3L–PARP9 co-evolve as a single adaptive unit linking ubiquitination and ADP-ribosylation in antiviral immunity

doi: 10.64898/2026.04.23.720386

Figure Lengend Snippet: Phylogenetic trees of bats (left) and rodent (right) DTX3L, PARP9 and PARP14, showing similar branches and nodes under significant positive selection (p-value <0.05, in red).

Article Snippet: This pattern is noteworthy given that, within the DELTEX family, DTX3L and DTX3 are the only members harboring ssNA-ubiquitinating activity ( ).

Techniques: Selection

Journal: bioRxiv

Article Title: DTX3L–PARP9 co-evolve as a single adaptive unit linking ubiquitination and ADP-ribosylation in antiviral immunity

doi: 10.64898/2026.04.23.720386

Figure Lengend Snippet: a) Haplotype diversity across the DTX-PARP gene family at shorter evolutionary timescales. Haplotype diversity (Hd = N/(N−1) × (1 − Σpᵢ²), where pᵢ is the frequency of haplotype i and N the number of samples; ) computed for each gene across datasets spanning different levels of divergence, from populations within a species to subspecies and closely related species within a genus. Each bar represents one dataset (population or lineage), colored and grouped by genus. The top row shows the non-complex DTX paralogs (DTX2, DTX3, DTX4) and the bottom row shows the three members of the DTX3L-PARP9-PARP14 functional complex (highlighted with a blue background). b) Geographic distribution of amino-acid haplotypes in Chlorocebus (Africa). Pie charts are positioned at the mean geographic coordinates of each sampled African lineage and show the relative frequency of each amino-acid haplotype. Each label indicates the Chlorocebus species or subspecies following the taxonomy of . Colours are gene-specific and are ranked by global frequency across the genus (Haplotype 1 = most frequent). Caribbean-origin populations (Barbados, Saint Kitts, Nevis) are excluded. Haplotypes observed in fewer than three individuals per dataset are pooled as ’rare’ (black). Panels from left to right: DTX3L, PARP9, PARP14. c) Distribution of positively selected sites (PSS) and non-synonymous single nucleotide polymorphisms (SNPs) across the coding regions of DTX3L, PARP9, and PARP14 for human and cattle. For each gene, PSS are shown along the top track in grey, and non-synonymous SNPs are shown along the bottom track in yellow. Sites that are both PSS and SNPs are highlighted in red, indicating positions where long-term adaptive evolution and ongoing intraspecific variation coincide. d-e) Predicted 3D docking structures of DTX3L-PARP9 complex from human (d) and cattle (e) using Colabfold (pLDDT>70), showing the spatial conformation of DTX3L RING-DTC and PARP9 macrodomain and the positions of the non-synonymous polymorphisms (blue) compared to the positively selected sites (red).

Article Snippet: This pattern is noteworthy given that, within the DELTEX family, DTX3L and DTX3 are the only members harboring ssNA-ubiquitinating activity ( ).

Techniques: Functional Assay

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: Identification of USP10 peptide ( a ) and DTX3L peptide ( b ) in the SATB2–interacting proteins from H2S GSCs overexpressing SATB2-Myc by mass spectrometry analysis. c,d , CoIP assays of interaction between exogenous Flag-tagged USP10 and Myc-tagged SATB2 in HEK293T cells. Immunoblots of precipitated proteins or total lysates were performed using indicated antibodies. e , f CoIP assays of interaction between exogenous Flag-tagged DTX3L and Myc-tagged SATB2 in HEK293T cells. g CoIP assays of endogenous protein interaction in GSCs. Schematic diagram of Myc-tagged full-length (FL) SATB2 ( h ), Flag-tagged FL USP10 ( i ), Flag-tagged FL DTX3L ( j ), and their truncated mutants. k HEK293T cells were co-transfected with USP10-Flag and Myc-tagged FL SATB2 or its truncated mutants, and cell lysates were analyzed by IP with Myc antibody followed by IB with indicated antibodies. l HEK293T cells were co-transfected with SATB2-Myc and Flag-tagged FL USP10 or its truncated mutants, and cell lysates were analyzed by IP with Flag antibody followed by IB with indicated antibodies. m HEK293T cells were co-transfected with DTX3L-Flag and Myc-tagged FL SATB2 or its truncated mutants, and cell lysates were analyzed by IP with Myc antibody followed by IB with indicated antibodies. n HEK293T cells were co-transfected with SATB2-Myc and Flag-tagged FL DTX3L or its truncated mutants, and cell lysates were analyzed by IP with Flag antibody followed by IB with indicated antibodies. Independent experiments were repeated three times, with similar results ( c – g , k – n ).

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: Mass Spectrometry, Western Blot, Transfection

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 USP10, DTX3L, and SATB2 expression in cell lysates of GSCs and matched NSTCs. qPCR analysis of USP10 ( b ) or DTX3L ( c ) mRNA expression in GSCs and matched NSTCs. n = 3 biological independent samples. d qPCR analysis of USP10 and DTX3L mRNA expression in GSCs. n = 3 biological independent samples. e Immunofluorescence of USP10 (green) and SATB2 (red) in GSCs. Scale bar, 20 μm. f Immunofluorescence of DTX3L (green) and SATB2 (red) in GSCs. Scale bar, 20 μm. g Immunofluorescence of USP10 (green) and the GSC marker SOX2 (red) in human GBM surgical specimens. Scale bar, 40 μm. h Immunofluorescence of DTX3L (green) and the GSC marker SOX2 (red) in human GBM specimens. Scale bar, 40 μm. i Immunofluorescence of USP10 (green) and SATB2 (red) in human GBM surgical specimens. Scale bar, 40 μm. j Immunofluorescence of DTX3L (green) and SATB2 (red) in human GBM specimens. Scale bar, 40 μm. Data are represented as mean ± s.d. Two-tailed unpaired t -test ( b – d ). Independent experiments were repeated three times, with similar results ( a, e – j ). 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, Immunofluorescence, Marker, Two Tailed Test

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 and DTX3L expression in GSCs transduced with vector control or DTX3L-Flag overexpression. b qPCR analysis of SATB2 mRNA expression in GSCs transduced with vector control or DTX3L-Flag overexpression. n = 3 biological independent samples. c Cell viability assay of GSCs transduced with vector control or DTX3L-Flag overexpression. n = 4 biological independent samples. d EdU incorporation assay of GSCs transduced with vector control or DTX3L-Flag overexpression. Scale bar, 45 μm. e Quantification of the ratio of EdU + cells in GSCs expressing vector control or DTX3L-Flag. n = 6 biological independent samples. f Tumorsphere images of GSCs transduced with vector control or DTX3L-Flag overexpression. Scale bar, 100 μm. Quantification of the number ( g ) and diameter ( h ) of tumorspheres formed by GSCs expressing vector control or DTX3L-Flag. n = 4 biological independent cell cultures. i In vitro limiting dilution analysis of the tumorsphere formations of GSCs transduced with vector control or DTX3L-Flag overexpression. j Kaplan-Meier survival curves of mice intracranially implanted with GSCs expressing vector control or DTX3L-Flag. n = 5–6 mice. k Representative H&E images of mouse brains collected at day 28 (T387GSCs) or 22 (H2S GSCs) after intracranially implantation with GSCs expressing vector control or DTX3L-Flag overexpression. Scale bar, 2 mm. Immunofluorescence of Ki67 (green) ( l ) and quantification of Ki67 + cells ( m ) in tumor xenografts derived from GSCs expressing vector control or DTX3L-Flag. Scale bar, 50 μm. n = 5 tumors. Immunofluorescence of SOX2 (red) ( n ) and quantification of SOX2 + cells ( o ) in tumor xenografts derived from GSCs expressing vector control or DTX3L-Flag. Scale bar, 50 μm. n = 5 tumors. Data are represented as mean ± s.d. Two-tailed unpaired t -test ( b , e , g , h , m , and o ), two-way ANOVA analysis followed by Sidak’s test ( c ), ELDA analysis (two-sided) for differences in stem cell frequencies ( i ), and log-rank test ( j ). Independent experiments were repeated three times, with similar results ( a and k ). 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, Control, Over Expression, Viability Assay, In Vitro, Immunofluorescence, Derivative Assay, Two Tailed Test

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: GSCs expressing DTX3L-Flag (T387, a , or H2S, b ) or shUSP10-1(T387, c , or H2S, d) along with their corresponding controls, were treated with CHX (100 μg/ml) and harvested at the indicated times for IB analysis. GSCs expressing DTX3L-Flag ( e ) or shUSP10 ( f ), along with their corresponding controls, were treated with MG-132 (20 µM) or vehicle control for 12 hours and then harvested for IB analysis. g HEK293T cells were co-transfected with SATB2-Myc, DTX3L-Flag, and His-Ub, and then cell lysates were subjected to IP with Myc antibody followed by IB with indicated antibodies. h GSCs were transduced with vector control or DTX3L-Flag, and then cell lysates were subjected to IP with SATB2 antibody or IgG followed by IB with ubiquitin (Ub) antibody. i HEK293T cells were co-transfected with SATB2-Myc, USP10-Flag, and His-Ub, and then cell lysates were subjected to IP with Myc antibody followed by IB with His antibody. j GSCs were transduced with shNT or shUSP10, and then cell lysates were subjected to IP with SATB2 antibody or IgG, followed by IB with Ub antibody. k HEK293T cells were co-transfected with SATB2-Myc, DTX3L-Flag, and the indicated His-tagged WT Ub, Lys48-only (K48) Ub, or Lys63-only (K63) Ub. Thereafter, cell lysates were subjected to IP with Myc antibody followed by IB with the indicated antibodies. l HEK293T cells were co-transfected with SATB2-Myc, DTX3L-Flag, and the indicated His-tagged WT Ub, Lys6-only (K6) Ub, Lys11-only (K11) Ub, Lys27-only (K27) Ub, Lys29-only (K29) Ub, or Lys33-only (K33) Ub. Thereafter, cell lysates were subjected to IP with Myc antibody followed by IB with His antibody. m HEK293T cells were co-transfected with SATB2-Myc, USP10-Flag, and the indicated His-tagged WT Ub, K48 Ub, or K63 Ub. Thereafter, cell lysates were subjected to IP with Myc antibody followed by IB with His antibody. n HEK293T cells were co-transfected with SATB2-Myc, USP10-Flag, and the indicated His-tagged WT Ub, K6 Ub, K11 Ub, K27 Ub, K29 Ub, or K33 Ub. Thereafter, cell lysates were subjected to IP with Myc antibody followed by IB with His antibody. Independent experiments were repeated three times, with similar results ( a – n ).

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, Control, Transfection, Transduction, Plasmid Preparation, Ubiquitin Proteomics

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

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 HEK293T cells were co-transfected with USP10-Flag and Myc-tagged truncated mutants of SATB2, and then cell lysates were analyzed by IB. b HEK293T cells were co-transfected with DTX3L-Flag and Myc-tagged truncated mutants of SATB2, and then cell lysates were analyzed by IB. c HEK293T cells were co-transfected with USP10-Flag and different lysine mutants of Myc-tagged truncated SATB2 (amino acids 232-437), and then cell lysates were analyzed by IB. d HEK293T cells were co-transfected with DTX3L-Flag and different lysine mutants of Myc-tagged truncated SATB2 (amino acids 232-437), and then cell lysates were analyzed by IB. e Alignment of the amino acid sequences of SATB2 peptides corresponding to human K266 from six species. f Mass spectrometry analysis of the peptide derived from SATB2 revealed ubiquitin conjugation at K266. HEK293T cells were transfected with SATB2-Myc and His-Ub constructs. Cell lysates were immunoprecipitated with anti-Myc antibody, and the captured SATB2-Myc was analyzed by mass spectrometry to identify ubiquitination sites. IB analysis of Myc and USP10 expression in GSCs (T387, g , or H2S, h ) transduced with Myc-tagged full-length SATB2-WT or SATB2-K266R in combination with shNT or shUSP10. IB analysis of Myc and Flag expression in GSCs (T387, i , or H2S, j ) transduced with Myc-tagged full-length of SATB2-WT or SATB2-K266R in combination with Flag-tagged DTX3L or vector control. k HEK293T cells were co-transfected with DTX3L-Flag, His-Ub, and Myc-tagged SATB2 WT or K266R (KR) mutant, and then cell lysates were subjected to IP with Myc antibody followed by IB with His antibody. l , HEK293T cells were co-transfected with USP10-Flag, His-Ub, and Myc-tagged SATB2 WT or K266R (KR) mutant, and cell then lysates were subjected to IP with Myc antibody followed by IB with His antibody. Independent experiments were repeated three times, with similar results ( a – d, g – l ).

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: Transfection, Mass Spectrometry, Derivative Assay, Ubiquitin Proteomics, Conjugation Assay, Construct, Immunoprecipitation, Expressing, Transduction, Plasmid Preparation, Control, Mutagenesis

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: USP10 antagonizes DTX3L-mediated ubiquitination at the K266 residue, thereby stabilizing the SATB2 protein, which in turn contributes to the malignant phenotype of GSCs and the growth of GBM tumor. Pharmacological inhibition of USP10 by Wu-5 promotes SATB2 ubiquitination and consequently suppresses GSC proliferation, self-renewal, and tumor growth. Created in BioRender. Guo, M. (2025) https://BioRender.com/p68d437 .

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: Ubiquitin Proteomics, Residue, Inhibition

Journal: Scientific Reports

Article Title: Pan-cancer analysis of DTX3L as a potential prognostic and immunological biomarker

doi: 10.1038/s41598-025-32552-3

Figure Lengend Snippet: The expression level of DTX3L in pan-cancer. ( A ) The expression level of DTX3L mRNA in pan-cancer. ( B ) The expression level of DTX3L protein in various tumors. Statistical method for ( A ) is Unpaired Wilcoxon Rank Sum and Signed Rank Test, for ( B ) is Mann-Whitney U test. Z-values represent standard deviations from the median across samples for the given cancer type. * P < 0.05; ** P < 0.01; **** P < 0.0001.

Article Snippet: Antibodies used included: DTX3L (1:1000, Proteintech Group, Wuhan, China), GAPDH (1:10000, Abcam, UK), and HRP-Goat anti Rabbit (1:10000, Abcam, UK).

Techniques: Expressing, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Pan-cancer analysis of DTX3L as a potential prognostic and immunological biomarker

doi: 10.1038/s41598-025-32552-3

Figure Lengend Snippet: Prognostic values of DTX3L in pan-cancer. ( A ) Correlation between DTX3L expression and OS. ( B ) PFS. ( C ) DSS. ( D ) DFS. Statistical methods: Cox regression analysis of the association between DTX3L expression and prognosis in each tumor type; log-rank test was used to determine prognostic significance. Hazard ratios (HR) and 95% confidence intervals (CI) are shown.

Article Snippet: Antibodies used included: DTX3L (1:1000, Proteintech Group, Wuhan, China), GAPDH (1:10000, Abcam, UK), and HRP-Goat anti Rabbit (1:10000, Abcam, UK).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Pan-cancer analysis of DTX3L as a potential prognostic and immunological biomarker

doi: 10.1038/s41598-025-32552-3

Figure Lengend Snippet: Correlation of DTX3L expression with clinical characteristics and RNA modification in pan-cancer. ( A ) T/N/M stages. ( B ) Tumor grade. ( C ) Tumor stage. ( D ) Patient age. ( E ) Correlation analysis of RNA gene modifications with DTX3L. Statistical methods: The unpaired Student’s t-test for comparisons between two groups and by one-way analysis of variance (ANOVA) for comparisons across multiple groups. The correlation coefficient (r) and p-value were calculated using Pearson correlation analysis. * P < 0.05; ** P < 0.01; *** P < 0.001, **** P < 0.0001.

Article Snippet: Antibodies used included: DTX3L (1:1000, Proteintech Group, Wuhan, China), GAPDH (1:10000, Abcam, UK), and HRP-Goat anti Rabbit (1:10000, Abcam, UK).

Techniques: Expressing, RNA modification

Journal: Scientific Reports

Article Title: Pan-cancer analysis of DTX3L as a potential prognostic and immunological biomarker

doi: 10.1038/s41598-025-32552-3

Figure Lengend Snippet: The correlation between DTX3L expression and immune feature in pan-cancer. ( A ) The correlation between DTX3L expression and immune checkpoint genes. ( B ) The correlation between DTX3L expression and immune cells. ( C ) The correlation between DTX3L expression and immune infiltration scores. Statistical method: The correlation coefficient (r) and p-value were calculated using Pearson correlation analysis.

Article Snippet: Antibodies used included: DTX3L (1:1000, Proteintech Group, Wuhan, China), GAPDH (1:10000, Abcam, UK), and HRP-Goat anti Rabbit (1:10000, Abcam, UK).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Pan-cancer analysis of DTX3L as a potential prognostic and immunological biomarker

doi: 10.1038/s41598-025-32552-3

Figure Lengend Snippet: Functional enrichment of DTX3L-related genes in pan-cancer. ( A ) DTX3L-related genes are shown in a network generated with the STRING website. ( B ) GO-biological process. ( C ) GO-cellular component. ( D ) GO-molecular function. (E) KEGG Pathways.

Article Snippet: Antibodies used included: DTX3L (1:1000, Proteintech Group, Wuhan, China), GAPDH (1:10000, Abcam, UK), and HRP-Goat anti Rabbit (1:10000, Abcam, UK).

Techniques: Functional Assay, Generated

Journal: Scientific Reports

Article Title: Pan-cancer analysis of DTX3L as a potential prognostic and immunological biomarker

doi: 10.1038/s41598-025-32552-3

Figure Lengend Snippet: DTX3L overexpression in OV and knockdown inhibits cell proliferation/migration in vitro. ( A ) DTX3L expression in OV tissues and normal tissues tested by IHC (×10). ( B ) DTX3L expression in ovarian cancer and normal ovarian cells. ( C ) Validation of DTX3L knockdown efficiency in A2780 cell. ( D ) CCK8 detection of cell proliferation. ( E ) Wound healing assay detection of cell migration. ( F ) Flow cytometry detection of cell apoptosis. Statistical method: Unpaired Student’s t-tests were performed to assess significant differences between two groups. Data are presented as mean ± SD. ** P < 0.01;* P < 0.05.

Article Snippet: Antibodies used included: DTX3L (1:1000, Proteintech Group, Wuhan, China), GAPDH (1:10000, Abcam, UK), and HRP-Goat anti Rabbit (1:10000, Abcam, UK).

Techniques: Over Expression, Knockdown, Migration, In Vitro, Expressing, Biomarker Discovery, Wound Healing Assay, Flow Cytometry