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Proteintech nono polyclonal antibody

DNA damage enhances the association between <t>NONO</t> and OGT. A Left panel: IR induces O-GlcNAcylation of NONO. HEK293T cells were exposed to 0, 2, 5 or 10 Gy of IR. Following treatment, cells were lysed using NETN300 buffer, and immunoprecipitation (IP) and Western blot were performed with the indicated antibodies. Right panel: Quantification of NONO O-GlcNAcylation levels, normalized to NONO levels and presented as fold change relative to control samples ( n = 3 per condition). B , C Investigation of the exogenous interaction between OGT and NONO via Co-IP. HEK293T cells were transfected with SFB-NONO and Myc-OGT plasmids. IP followed by Western blot was conducted using the indicated antibodies to assess the interaction between OGT and NONO. D Left panel: DNA damage enhances the endogenous interaction between NONO and OGT. HEK293T cells were exposed to 10 Gy of IR and lysed with NETN300 buffer. Cell extracts were subjected to IP and Western blot analysis using the indicated antibodies. Right panel: Quantification of OGT level, normalized to NONO levels and presented as fold change relative to control samples ( n = 3 per condition)

Nono Polyclonal Antibody, supplied by Proteintech, used in various techniques. Bioz Stars score: 95/100, based on 61 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/nono+polyclonal+antibody/pmc12888201-333-24-27?v=Proteintech
Average 95 stars, based on 61 article reviews

nono polyclonal antibody - by Bioz Stars, 2026-07

95/100 stars

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1) Product Images from "O-GlcNAcylation of NONO mediates alternative splicing of SETMAR and facilitates NHEJ repair"

Article Title: O-GlcNAcylation of NONO mediates alternative splicing of SETMAR and facilitates NHEJ repair

Journal: Genome Biology

doi: 10.1186/s13059-026-03930-5

DNA damage enhances the association between NONO and OGT. A Left panel: IR induces O-GlcNAcylation of NONO. HEK293T cells were exposed to 0, 2, 5 or 10 Gy of IR. Following treatment, cells were lysed using NETN300 buffer, and immunoprecipitation (IP) and Western blot were performed with the indicated antibodies. Right panel: Quantification of NONO O-GlcNAcylation levels, normalized to NONO levels and presented as fold change relative to control samples ( n = 3 per condition). B , C Investigation of the exogenous interaction between OGT and NONO via Co-IP. HEK293T cells were transfected with SFB-NONO and Myc-OGT plasmids. IP followed by Western blot was conducted using the indicated antibodies to assess the interaction between OGT and NONO. D Left panel: DNA damage enhances the endogenous interaction between NONO and OGT. HEK293T cells were exposed to 10 Gy of IR and lysed with NETN300 buffer. Cell extracts were subjected to IP and Western blot analysis using the indicated antibodies. Right panel: Quantification of OGT level, normalized to NONO levels and presented as fold change relative to control samples ( n = 3 per condition)

Figure Legend Snippet: DNA damage enhances the association between NONO and OGT. A Left panel: IR induces O-GlcNAcylation of NONO. HEK293T cells were exposed to 0, 2, 5 or 10 Gy of IR. Following treatment, cells were lysed using NETN300 buffer, and immunoprecipitation (IP) and Western blot were performed with the indicated antibodies. Right panel: Quantification of NONO O-GlcNAcylation levels, normalized to NONO levels and presented as fold change relative to control samples ( n = 3 per condition). B , C Investigation of the exogenous interaction between OGT and NONO via Co-IP. HEK293T cells were transfected with SFB-NONO and Myc-OGT plasmids. IP followed by Western blot was conducted using the indicated antibodies to assess the interaction between OGT and NONO. D Left panel: DNA damage enhances the endogenous interaction between NONO and OGT. HEK293T cells were exposed to 10 Gy of IR and lysed with NETN300 buffer. Cell extracts were subjected to IP and Western blot analysis using the indicated antibodies. Right panel: Quantification of OGT level, normalized to NONO levels and presented as fold change relative to control samples ( n = 3 per condition)

Techniques Used: Immunoprecipitation, Western Blot, Control, Co-Immunoprecipitation Assay, Transfection

$O-GlcNAcylation enhances the stability of NONO protein. A Left panel: O-GlcNAcylation promotes NONO recruitment to DNA damage sites. U2OS cells expressing GFP-NONO were subjected to laser microirradiation, with or without OSMI-1 treatment. Scale bar: 5 μm. Right panel: Quantification of GFP-NONO intensity at laser-induced damage sites. The intensity of GFP-NONO at laser stripes was quantified at indicated time points using Image J, and peak fluorescence density in the micro-irradiated areas was plotted against time. B Left panel: Inhibition of O-GlcNAcylation reduces NONO binding to chromatin. HEK293T cells were transfected with SFB-NONO and treated with either DMSO or 20 μM OSMI-1 for 48 h. Whole cell extract (WCE), non-chromatin, and chromatin fractions of cells were harvested and analyzed by immunoblotting with the indicated antibodies. Right panel: Quantification of Flag (NONO) level, normalized to histone H3 levels in chromatin and presented as fold change relative to control samples ( n = 3 per condition). C Left panel: Time-dependent degradation of NONO upon OGT inhibition. HEK293T cells were treated with 20 μM OSMI-1 or mock treatment for up to 72 h. NONO protein levels were assessed using Western blot, with GAPDH serving as the loading control. Right panel: Quantification of NONO level, normalized to GAPDH levels and presented as fold change relative to control samples ( n = 3 per condition). D Left panel: Inhibition of O-GlcNAcylation accelerates NONO degradation. HEK293T cells were treated with either DMSO or 20 μM OSMI-1 for 48 h, followed by 50 μM CHX treatment for up to 8 h. NONO protein levels were monitored using Western blot analysis, with GAPDH as the loading control. Right panel: Quantification of NONO level, normalized to GAPDH and presented as fold change relative to control samples ( n = 3 per condition)$
Figure Legend Snippet: O-GlcNAcylation enhances the stability of NONO protein. A Left panel: O-GlcNAcylation promotes NONO recruitment to DNA damage sites. U2OS cells expressing GFP-NONO were subjected to laser microirradiation, with or without OSMI-1 treatment. Scale bar: 5 μm. Right panel: Quantification of GFP-NONO intensity at laser-induced damage sites. The intensity of GFP-NONO at laser stripes was quantified at indicated time points using Image J, and peak fluorescence density in the micro-irradiated areas was plotted against time. B Left panel: Inhibition of O-GlcNAcylation reduces NONO binding to chromatin. HEK293T cells were transfected with SFB-NONO and treated with either DMSO or 20 μM OSMI-1 for 48 h. Whole cell extract (WCE), non-chromatin, and chromatin fractions of cells were harvested and analyzed by immunoblotting with the indicated antibodies. Right panel: Quantification of Flag (NONO) level, normalized to histone H3 levels in chromatin and presented as fold change relative to control samples ( n = 3 per condition). C Left panel: Time-dependent degradation of NONO upon OGT inhibition. HEK293T cells were treated with 20 μM OSMI-1 or mock treatment for up to 72 h. NONO protein levels were assessed using Western blot, with GAPDH serving as the loading control. Right panel: Quantification of NONO level, normalized to GAPDH levels and presented as fold change relative to control samples ( n = 3 per condition). D Left panel: Inhibition of O-GlcNAcylation accelerates NONO degradation. HEK293T cells were treated with either DMSO or 20 μM OSMI-1 for 48 h, followed by 50 μM CHX treatment for up to 8 h. NONO protein levels were monitored using Western blot analysis, with GAPDH as the loading control. Right panel: Quantification of NONO level, normalized to GAPDH and presented as fold change relative to control samples ( n = 3 per condition)

Techniques Used: Expressing, Fluorescence, Irradiation, Inhibition, Binding Assay, Transfection, Western Blot, Control

$O-GlcNAcylation at S147 is required for NONO-mediated DNA damage repair. A Assessment of NONO O-GlcNAcylation. HEK293T cells were transfected with empty vector (EV), SFB-NONO-WT, S147A, T440A or 2 A plasmids, then treated by 10 Gy of IR. Cell lysates were denatured, immunoprecipitated with streptavidin beads, and analyzed via immunoblotting with O-GlcNAc and Flag antibodies. B Conservation of the S147 locus across different species. The upper panel depicts wild-type NONO, including RNA recognition motifs (RRM1/2), the NOPS domain, coiled-coil domain (CC), and C-terminal intrinsically disordered domains. The lower panel shows a BLAST alignment of the S147 locus across species. C Left panel: O-GlcNAcylation of Ser147 facilitates NONO recruitment to DNA damage sites. U2OS cells transfected with GFP-NONO-WT or S147A were subjected to laser microirradiation. Scale bar: 5 μm. Right panel: Quantification of GFP-NONO intensity at laser strips using Image J, and peak fluorescence density in the micro-irradiated areas was plotted against time. D Left panel: S147A exhibited impaired chromatin-binding without MG132 treatment. HEK293T cells transfected with SFB-NONO-WT or S147A were subjected to fraction extraction and immunoblotting. Right panel: Quantification of Flag (NONO) levels ( n = 3 per condition). E Left panel: O-GlcNAcylation of S147 is essential for NONO-mediated DNA damage repair. Stable HEK293T cells with NONO knockdown and reconstituted WT or S147A were exposed to 10 Gy of IR and underwent neutral comet assays (* P < 0.05). Right panel: Quantitative analysis of comet assay results from three independent experiments (50 cells per time point). Data are presented as mean ± SEM. (F) O-GlcNAcylation of Ser147 promotes NONO-mediated NHEJ repair. NONO knockdown HEK293T cells reconstituted WT or S147A were transfected with an NHEJ reporter pre-digested by Hind III and cultured for 48 h. FACS analysis quantified GFP-positive cells. Data from three independent experiments are presented as mean ± SD. ** P < 0.01, *** P < 0.001. Stable HEK293T cells with Ku80 knockdown were used as a positive control$
Figure Legend Snippet: O-GlcNAcylation at S147 is required for NONO-mediated DNA damage repair. A Assessment of NONO O-GlcNAcylation. HEK293T cells were transfected with empty vector (EV), SFB-NONO-WT, S147A, T440A or 2 A plasmids, then treated by 10 Gy of IR. Cell lysates were denatured, immunoprecipitated with streptavidin beads, and analyzed via immunoblotting with O-GlcNAc and Flag antibodies. B Conservation of the S147 locus across different species. The upper panel depicts wild-type NONO, including RNA recognition motifs (RRM1/2), the NOPS domain, coiled-coil domain (CC), and C-terminal intrinsically disordered domains. The lower panel shows a BLAST alignment of the S147 locus across species. C Left panel: O-GlcNAcylation of Ser147 facilitates NONO recruitment to DNA damage sites. U2OS cells transfected with GFP-NONO-WT or S147A were subjected to laser microirradiation. Scale bar: 5 μm. Right panel: Quantification of GFP-NONO intensity at laser strips using Image J, and peak fluorescence density in the micro-irradiated areas was plotted against time. D Left panel: S147A exhibited impaired chromatin-binding without MG132 treatment. HEK293T cells transfected with SFB-NONO-WT or S147A were subjected to fraction extraction and immunoblotting. Right panel: Quantification of Flag (NONO) levels ( n = 3 per condition). E Left panel: O-GlcNAcylation of S147 is essential for NONO-mediated DNA damage repair. Stable HEK293T cells with NONO knockdown and reconstituted WT or S147A were exposed to 10 Gy of IR and underwent neutral comet assays (* P < 0.05). Right panel: Quantitative analysis of comet assay results from three independent experiments (50 cells per time point). Data are presented as mean ± SEM. (F) O-GlcNAcylation of Ser147 promotes NONO-mediated NHEJ repair. NONO knockdown HEK293T cells reconstituted WT or S147A were transfected with an NHEJ reporter pre-digested by Hind III and cultured for 48 h. FACS analysis quantified GFP-positive cells. Data from three independent experiments are presented as mean ± SD. ** P < 0.01, *** P < 0.001. Stable HEK293T cells with Ku80 knockdown were used as a positive control

Techniques Used: Transfection, Plasmid Preparation, Immunoprecipitation, Western Blot, Fluorescence, Irradiation, Binding Assay, Extraction, Knockdown, Single Cell Gel Electrophoresis, Cell Culture, Positive Control

Ser147 O-GlcNAcylation of NONO antagonizes its ubiquitination by decreasing interaction with RNF8. A The proteasome inhibitor MG132 prevents degradation of the NONO S147A mutant. HEK293T cells were transfected with SFB-NONO-WT or the S147A mutant and treated with or without 10 μM MG132. Cell lysates were analyzed by Western blot analysis with the indicated antibodies. B Left panel: O-GlcNAcylation at S147 is critical for NONO stability. HEK293T cells expressing SFB-NONO-WT or S147A were treated with 50 μM CHX for 12 h. Protein levels were monitored by Western blot using anti-FLAG antibodies, with GAPDH serving as the loading control. Right panel: Quantification of Flag (NONO) expression, normalized to GAPDH levels and presented as fold change relative to control samples ( n = 3 per condition). C O-GlcNAcylation of NONO crosstalks with ubiquitination. HEK293T cells were transfected with Myc-RNF8, HA-Ub, and either SFB-NONO WT or the S147A mutant, followed by an in vivo ubiquitination assay. D O-GlcNAcylation of Ser147 impairs the interaction between NONO and RNF8. HEK293T cells were transfected with Myc-RNF8 and either SFB-NONO WT or S147A mutant. The interaction was assessed by IP assay

Figure Legend Snippet: Ser147 O-GlcNAcylation of NONO antagonizes its ubiquitination by decreasing interaction with RNF8. A The proteasome inhibitor MG132 prevents degradation of the NONO S147A mutant. HEK293T cells were transfected with SFB-NONO-WT or the S147A mutant and treated with or without 10 μM MG132. Cell lysates were analyzed by Western blot analysis with the indicated antibodies. B Left panel: O-GlcNAcylation at S147 is critical for NONO stability. HEK293T cells expressing SFB-NONO-WT or S147A were treated with 50 μM CHX for 12 h. Protein levels were monitored by Western blot using anti-FLAG antibodies, with GAPDH serving as the loading control. Right panel: Quantification of Flag (NONO) expression, normalized to GAPDH levels and presented as fold change relative to control samples ( n = 3 per condition). C O-GlcNAcylation of NONO crosstalks with ubiquitination. HEK293T cells were transfected with Myc-RNF8, HA-Ub, and either SFB-NONO WT or the S147A mutant, followed by an in vivo ubiquitination assay. D O-GlcNAcylation of Ser147 impairs the interaction between NONO and RNF8. HEK293T cells were transfected with Myc-RNF8 and either SFB-NONO WT or S147A mutant. The interaction was assessed by IP assay

Techniques Used: Ubiquitin Proteomics, Mutagenesis, Transfection, Western Blot, Expressing, Control, In Vivo

O-GlcNAcylation of NONO promotes NHEJ by regulating the alternative splicing of pre-mRNA of SETMAR. A Schematic illustrating how the NONO-SFPQ complex influences NHEJ repair by modulating the alternative splicing of SETMAR precursor mRNA. B O-GlcNAcylation of Ser147 stabilizes the assembly of the NONO-SFPQ complex. HEK293T cells were transfected with either SFB-NONO or the S147A mutant. Cell lysates were immunoprecipitated with Streptavidin beads, followed by Western blotting with anti-SFPQ or anti-Flag antibodies. C Quantification of qPCR analysis of SETMAR pre-mRNA from the RIP assay of HEK293T cells. Cells were transfected with SFB-NONO, S147A mutant, or an empty vector. Cell lysates were subjected to RIP, and the products were amplified by qPCR using the indicated primers pairs. Input was used for normalization, and the empty vector served as a negative control. D Western blot analysis of RNA pull-down eluates using anti-Myc antibodies demonstrates a higher binding specificity of NONO-WT for CAGGCAGG RNA repeats compared to the S147A mutant in HEK293T cells. E Quantification of the SETMAR-L/SETMAR-S ratio in HEK293T cells by qPCR. F Left panel: RNA FISH analysis shows that O-GlcNAcylation of NONO promotes the expression of SETMAR-L. RNA FISH was conducted to visualize SETMAR-L (green) and SETMAR-S (red) in NONO-knockdown HEK293T cells stably reconstituted with either WT NONO or the S147A mutant. DAPI staining of the nuclei is shown in blue. Scale bar, 20 μm. Right panel: Quantification of fluorescence intensity from FISH. Data from three independent experiments are presented as mean ± SD. Scale bar, 20 μm

Figure Legend Snippet: O-GlcNAcylation of NONO promotes NHEJ by regulating the alternative splicing of pre-mRNA of SETMAR. A Schematic illustrating how the NONO-SFPQ complex influences NHEJ repair by modulating the alternative splicing of SETMAR precursor mRNA. B O-GlcNAcylation of Ser147 stabilizes the assembly of the NONO-SFPQ complex. HEK293T cells were transfected with either SFB-NONO or the S147A mutant. Cell lysates were immunoprecipitated with Streptavidin beads, followed by Western blotting with anti-SFPQ or anti-Flag antibodies. C Quantification of qPCR analysis of SETMAR pre-mRNA from the RIP assay of HEK293T cells. Cells were transfected with SFB-NONO, S147A mutant, or an empty vector. Cell lysates were subjected to RIP, and the products were amplified by qPCR using the indicated primers pairs. Input was used for normalization, and the empty vector served as a negative control. D Western blot analysis of RNA pull-down eluates using anti-Myc antibodies demonstrates a higher binding specificity of NONO-WT for CAGGCAGG RNA repeats compared to the S147A mutant in HEK293T cells. E Quantification of the SETMAR-L/SETMAR-S ratio in HEK293T cells by qPCR. F Left panel: RNA FISH analysis shows that O-GlcNAcylation of NONO promotes the expression of SETMAR-L. RNA FISH was conducted to visualize SETMAR-L (green) and SETMAR-S (red) in NONO-knockdown HEK293T cells stably reconstituted with either WT NONO or the S147A mutant. DAPI staining of the nuclei is shown in blue. Scale bar, 20 μm. Right panel: Quantification of fluorescence intensity from FISH. Data from three independent experiments are presented as mean ± SD. Scale bar, 20 μm

Techniques Used: Alternative Splicing, Transfection, Mutagenesis, Immunoprecipitation, Western Blot, Plasmid Preparation, Amplification, Negative Control, Binding Assay, Expressing, Knockdown, Stable Transfection, Staining, Fluorescence

$O-GlcNAcylation of NONO facilitates H3K36me2 enrichment at DSBs and recruits Ku70 for NHEJ. A Left panel: O-GlcNAcylation at Ser147 stabilizes Ku70 recruitment at DNA damage sites. U2OS cells were transfected with GFP-Ku70, along with an empty vector, SFB-NONO WT, or the S147A mutant. Representative images demonstrated the dynamic recruitment of GFP-Ku70 to DNA damage sites. Data were presented as mean ± SEM. Scale bar, 10 μm. Right panel: Quantification of the time course of GFP-Ku70 recruitment following laser microirradiation. B Upper panel: O-GlcNAcylation at Ser147 stabilizes the binding of Ku70 to chromatin. HEK293T cells were transfected with an empty vector, SFB-NONO WT, or the S147A mutant and exposed to 10 Gy of IR. Following IR, cells were allowed to recover for different durations. Whole cell extract (WCE) and chromatin fractions were prepared and subjected to IP and Western blot analysis with the indicated antibodies. Lower panel: Quantification of Ku70 level, normalized to H3 levels in chromatin and presented as fold change relative to control samples ( n = 3 per condition). C Upper panel: Impact of O-GlcNAcylation at Ser147 on H3K36 demethylation levels. Stable HEK293T cells with NONO knockdown were reconstituted with shRNA-resistant NONO WT or S147A mutant. These cells were treated with IR and examined via western blot analysis with the indicated antibodies to assess H3K36me2 levels. Lower panel: Quantification of H3K36me2 level, normalized to H3 levels in chromatin and presented as fold change relative to control samples ( n = 3 per condition)$
Figure Legend Snippet: O-GlcNAcylation of NONO facilitates H3K36me2 enrichment at DSBs and recruits Ku70 for NHEJ. A Left panel: O-GlcNAcylation at Ser147 stabilizes Ku70 recruitment at DNA damage sites. U2OS cells were transfected with GFP-Ku70, along with an empty vector, SFB-NONO WT, or the S147A mutant. Representative images demonstrated the dynamic recruitment of GFP-Ku70 to DNA damage sites. Data were presented as mean ± SEM. Scale bar, 10 μm. Right panel: Quantification of the time course of GFP-Ku70 recruitment following laser microirradiation. B Upper panel: O-GlcNAcylation at Ser147 stabilizes the binding of Ku70 to chromatin. HEK293T cells were transfected with an empty vector, SFB-NONO WT, or the S147A mutant and exposed to 10 Gy of IR. Following IR, cells were allowed to recover for different durations. Whole cell extract (WCE) and chromatin fractions were prepared and subjected to IP and Western blot analysis with the indicated antibodies. Lower panel: Quantification of Ku70 level, normalized to H3 levels in chromatin and presented as fold change relative to control samples ( n = 3 per condition). C Upper panel: Impact of O-GlcNAcylation at Ser147 on H3K36 demethylation levels. Stable HEK293T cells with NONO knockdown were reconstituted with shRNA-resistant NONO WT or S147A mutant. These cells were treated with IR and examined via western blot analysis with the indicated antibodies to assess H3K36me2 levels. Lower panel: Quantification of H3K36me2 level, normalized to H3 levels in chromatin and presented as fold change relative to control samples ( n = 3 per condition)

Techniques Used: Transfection, Plasmid Preparation, Mutagenesis, Binding Assay, Western Blot, Control, Knockdown, shRNA

O-GlcNAcylation of NONO promotes radioresistance in hepatocellular carcinoma. A Analysis of NONO O-GlcNAcylation was conducted in human liver epithelial cells (THLE2) and hepatocellular carcinoma cells (HepG2, HCCLM9, Huh7) usingIP/Western blotting with indicated antibodies. B Generation of stable HCCLM9 cells with NONO knockdown. Western blot analysis was performed to assess the efficiency of NONO knockdown using two shRNA constructs(shNONO #1 and shNONO #2). GAPDH was used as a loading control. C Upper panel: O-GlcNAcylation at Ser147 promotes cell survival post-IR treatment. NONO-knockdown HCCLM9 cells were transfected with SFB-NONO WT or S147A mutant and subjected to clonogenic survival assays post-IR treatment. Cells were treated with the indicated doses of IR and further incubated for 7–10 days. Lower panel: Quantitative analysis of clonogenic survival assays. D Schematic diagram illustrating the radiotherapy process for NTG mice. Mice injected with control or NONO knockdown cells, as well as cells reconstituted with NONO WT or S147A and wereexposed to 8 Gy of IR twice. E O-GlcNAcylation of NONO enhances radioresistance. Mice were subcutaneously injected with 7 × 10 6 control or NONO knockdown cells, or cells reconstituted with NONO WT or S147A, and exposed to 8 Gy of IR twice or not, when tumors reached a similar size (about 100 mm. 3 ). Representative images of xenograft tumors are shown ( n = 6/group). F Quantification analysis of xenograft tumor volumes from ( E ). Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA. ** P < 0.01, *** P < 0.001. G Hematoxylin and eosin (H&E) staining and IHC analysis of NONO and H3K36me2 in xenograft tumors were performed, comparing controland NONO knockdown groups, as well as tumors reconstituted with NONO WT or S147A ( n = 6). Scale bar, 30 μm. H Quantification of H3K36me2 levels from ( G ). Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA. ** P < 0.01, *** P < 0.001

Figure Legend Snippet: O-GlcNAcylation of NONO promotes radioresistance in hepatocellular carcinoma. A Analysis of NONO O-GlcNAcylation was conducted in human liver epithelial cells (THLE2) and hepatocellular carcinoma cells (HepG2, HCCLM9, Huh7) usingIP/Western blotting with indicated antibodies. B Generation of stable HCCLM9 cells with NONO knockdown. Western blot analysis was performed to assess the efficiency of NONO knockdown using two shRNA constructs(shNONO #1 and shNONO #2). GAPDH was used as a loading control. C Upper panel: O-GlcNAcylation at Ser147 promotes cell survival post-IR treatment. NONO-knockdown HCCLM9 cells were transfected with SFB-NONO WT or S147A mutant and subjected to clonogenic survival assays post-IR treatment. Cells were treated with the indicated doses of IR and further incubated for 7–10 days. Lower panel: Quantitative analysis of clonogenic survival assays. D Schematic diagram illustrating the radiotherapy process for NTG mice. Mice injected with control or NONO knockdown cells, as well as cells reconstituted with NONO WT or S147A and wereexposed to 8 Gy of IR twice. E O-GlcNAcylation of NONO enhances radioresistance. Mice were subcutaneously injected with 7 × 10 6 control or NONO knockdown cells, or cells reconstituted with NONO WT or S147A, and exposed to 8 Gy of IR twice or not, when tumors reached a similar size (about 100 mm. 3 ). Representative images of xenograft tumors are shown ( n = 6/group). F Quantification analysis of xenograft tumor volumes from ( E ). Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA. ** P < 0.01, *** P < 0.001. G Hematoxylin and eosin (H&E) staining and IHC analysis of NONO and H3K36me2 in xenograft tumors were performed, comparing controland NONO knockdown groups, as well as tumors reconstituted with NONO WT or S147A ( n = 6). Scale bar, 30 μm. H Quantification of H3K36me2 levels from ( G ). Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA. ** P < 0.01, *** P < 0.001

Techniques Used: Western Blot, Knockdown, shRNA, Construct, Control, Transfection, Mutagenesis, Incubation, Injection, Staining

Figure Legend Snippet: Proposed working model of NONO O-GlcNAcylation regulating NHEJ-mediated DNA damage repair

Techniques Used:

Image Search Results

Journal: Genome Biology

Article Title: O-GlcNAcylation of NONO mediates alternative splicing of SETMAR and facilitates NHEJ repair

doi: 10.1186/s13059-026-03930-5

Figure Lengend Snippet: DNA damage enhances the association between NONO and OGT. A Left panel: IR induces O-GlcNAcylation of NONO. HEK293T cells were exposed to 0, 2, 5 or 10 Gy of IR. Following treatment, cells were lysed using NETN300 buffer, and immunoprecipitation (IP) and Western blot were performed with the indicated antibodies. Right panel: Quantification of NONO O-GlcNAcylation levels, normalized to NONO levels and presented as fold change relative to control samples ( n = 3 per condition). B , C Investigation of the exogenous interaction between OGT and NONO via Co-IP. HEK293T cells were transfected with SFB-NONO and Myc-OGT plasmids. IP followed by Western blot was conducted using the indicated antibodies to assess the interaction between OGT and NONO. D Left panel: DNA damage enhances the endogenous interaction between NONO and OGT. HEK293T cells were exposed to 10 Gy of IR and lysed with NETN300 buffer. Cell extracts were subjected to IP and Western blot analysis using the indicated antibodies. Right panel: Quantification of OGT level, normalized to NONO levels and presented as fold change relative to control samples ( n = 3 per condition)

Article Snippet: Antibodies used in this study include the following: Anti-DDDDK-tag mAb (MBL, M185-3L), Anti-Myc-tag mAb (MBL, M047-3), Anti-GAPDH (Proteintech, 60,004–1-Ig), OGT polyclonal antibody (Proteintech, 11,576–2-AP), NONO polyclonal antibody (Proteintech, 11,058–1-AP), RL2 (Santa cruz biotechnology, SC-59624, which is an antibody against O-GlcNAc), Histone-H3 polyclonal antibody (Proteintech, 17,168–1-AP), anti-HA (Sigma, 66,006–2-Ig), Anti-PAR monoclonal antibody (R&D, 4335-MC-100-AC), Ku80 polyclonal antibody (Proteintech, 16,389–1-AP), SFPQ polyclonal antibody (Proteintech, 15,585–1-AP), anti-Histone H3(di-methyl K36) polyclonal antibody (abcam, AB9049), Ku70 polyclonal antibody (Proteintech, 10,723–1-AP), anti-SETMAR polyclonal antibody (abcam, ab129455), anti-RAD51 monoclonal antibody(abcam, ab133534).

Techniques: Immunoprecipitation, Western Blot, Control, Co-Immunoprecipitation Assay, Transfection

Journal: Genome Biology

Article Title: O-GlcNAcylation of NONO mediates alternative splicing of SETMAR and facilitates NHEJ repair

doi: 10.1186/s13059-026-03930-5

Figure Lengend Snippet: O-GlcNAcylation enhances the stability of NONO protein. A Left panel: O-GlcNAcylation promotes NONO recruitment to DNA damage sites. U2OS cells expressing GFP-NONO were subjected to laser microirradiation, with or without OSMI-1 treatment. Scale bar: 5 μm. Right panel: Quantification of GFP-NONO intensity at laser-induced damage sites. The intensity of GFP-NONO at laser stripes was quantified at indicated time points using Image J, and peak fluorescence density in the micro-irradiated areas was plotted against time. B Left panel: Inhibition of O-GlcNAcylation reduces NONO binding to chromatin. HEK293T cells were transfected with SFB-NONO and treated with either DMSO or 20 μM OSMI-1 for 48 h. Whole cell extract (WCE), non-chromatin, and chromatin fractions of cells were harvested and analyzed by immunoblotting with the indicated antibodies. Right panel: Quantification of Flag (NONO) level, normalized to histone H3 levels in chromatin and presented as fold change relative to control samples ( n = 3 per condition). C Left panel: Time-dependent degradation of NONO upon OGT inhibition. HEK293T cells were treated with 20 μM OSMI-1 or mock treatment for up to 72 h. NONO protein levels were assessed using Western blot, with GAPDH serving as the loading control. Right panel: Quantification of NONO level, normalized to GAPDH levels and presented as fold change relative to control samples ( n = 3 per condition). D Left panel: Inhibition of O-GlcNAcylation accelerates NONO degradation. HEK293T cells were treated with either DMSO or 20 μM OSMI-1 for 48 h, followed by 50 μM CHX treatment for up to 8 h. NONO protein levels were monitored using Western blot analysis, with GAPDH as the loading control. Right panel: Quantification of NONO level, normalized to GAPDH and presented as fold change relative to control samples ( n = 3 per condition)

Techniques: Expressing, Fluorescence, Irradiation, Inhibition, Binding Assay, Transfection, Western Blot, Control

Journal: Genome Biology

Article Title: O-GlcNAcylation of NONO mediates alternative splicing of SETMAR and facilitates NHEJ repair

doi: 10.1186/s13059-026-03930-5

Figure Lengend Snippet: O-GlcNAcylation at S147 is required for NONO-mediated DNA damage repair. A Assessment of NONO O-GlcNAcylation. HEK293T cells were transfected with empty vector (EV), SFB-NONO-WT, S147A, T440A or 2 A plasmids, then treated by 10 Gy of IR. Cell lysates were denatured, immunoprecipitated with streptavidin beads, and analyzed via immunoblotting with O-GlcNAc and Flag antibodies. B Conservation of the S147 locus across different species. The upper panel depicts wild-type NONO, including RNA recognition motifs (RRM1/2), the NOPS domain, coiled-coil domain (CC), and C-terminal intrinsically disordered domains. The lower panel shows a BLAST alignment of the S147 locus across species. C Left panel: O-GlcNAcylation of Ser147 facilitates NONO recruitment to DNA damage sites. U2OS cells transfected with GFP-NONO-WT or S147A were subjected to laser microirradiation. Scale bar: 5 μm. Right panel: Quantification of GFP-NONO intensity at laser strips using Image J, and peak fluorescence density in the micro-irradiated areas was plotted against time. D Left panel: S147A exhibited impaired chromatin-binding without MG132 treatment. HEK293T cells transfected with SFB-NONO-WT or S147A were subjected to fraction extraction and immunoblotting. Right panel: Quantification of Flag (NONO) levels ( n = 3 per condition). E Left panel: O-GlcNAcylation of S147 is essential for NONO-mediated DNA damage repair. Stable HEK293T cells with NONO knockdown and reconstituted WT or S147A were exposed to 10 Gy of IR and underwent neutral comet assays (* P < 0.05). Right panel: Quantitative analysis of comet assay results from three independent experiments (50 cells per time point). Data are presented as mean ± SEM. (F) O-GlcNAcylation of Ser147 promotes NONO-mediated NHEJ repair. NONO knockdown HEK293T cells reconstituted WT or S147A were transfected with an NHEJ reporter pre-digested by Hind III and cultured for 48 h. FACS analysis quantified GFP-positive cells. Data from three independent experiments are presented as mean ± SD. ** P < 0.01, *** P < 0.001. Stable HEK293T cells with Ku80 knockdown were used as a positive control

Techniques: Transfection, Plasmid Preparation, Immunoprecipitation, Western Blot, Fluorescence, Irradiation, Binding Assay, Extraction, Knockdown, Single Cell Gel Electrophoresis, Cell Culture, Positive Control

Journal: Genome Biology

Article Title: O-GlcNAcylation of NONO mediates alternative splicing of SETMAR and facilitates NHEJ repair

doi: 10.1186/s13059-026-03930-5

Figure Lengend Snippet: Ser147 O-GlcNAcylation of NONO antagonizes its ubiquitination by decreasing interaction with RNF8. A The proteasome inhibitor MG132 prevents degradation of the NONO S147A mutant. HEK293T cells were transfected with SFB-NONO-WT or the S147A mutant and treated with or without 10 μM MG132. Cell lysates were analyzed by Western blot analysis with the indicated antibodies. B Left panel: O-GlcNAcylation at S147 is critical for NONO stability. HEK293T cells expressing SFB-NONO-WT or S147A were treated with 50 μM CHX for 12 h. Protein levels were monitored by Western blot using anti-FLAG antibodies, with GAPDH serving as the loading control. Right panel: Quantification of Flag (NONO) expression, normalized to GAPDH levels and presented as fold change relative to control samples ( n = 3 per condition). C O-GlcNAcylation of NONO crosstalks with ubiquitination. HEK293T cells were transfected with Myc-RNF8, HA-Ub, and either SFB-NONO WT or the S147A mutant, followed by an in vivo ubiquitination assay. D O-GlcNAcylation of Ser147 impairs the interaction between NONO and RNF8. HEK293T cells were transfected with Myc-RNF8 and either SFB-NONO WT or S147A mutant. The interaction was assessed by IP assay

Techniques: Ubiquitin Proteomics, Mutagenesis, Transfection, Western Blot, Expressing, Control, In Vivo

Journal: Genome Biology

Article Title: O-GlcNAcylation of NONO mediates alternative splicing of SETMAR and facilitates NHEJ repair

doi: 10.1186/s13059-026-03930-5

Figure Lengend Snippet: O-GlcNAcylation of NONO promotes NHEJ by regulating the alternative splicing of pre-mRNA of SETMAR. A Schematic illustrating how the NONO-SFPQ complex influences NHEJ repair by modulating the alternative splicing of SETMAR precursor mRNA. B O-GlcNAcylation of Ser147 stabilizes the assembly of the NONO-SFPQ complex. HEK293T cells were transfected with either SFB-NONO or the S147A mutant. Cell lysates were immunoprecipitated with Streptavidin beads, followed by Western blotting with anti-SFPQ or anti-Flag antibodies. C Quantification of qPCR analysis of SETMAR pre-mRNA from the RIP assay of HEK293T cells. Cells were transfected with SFB-NONO, S147A mutant, or an empty vector. Cell lysates were subjected to RIP, and the products were amplified by qPCR using the indicated primers pairs. Input was used for normalization, and the empty vector served as a negative control. D Western blot analysis of RNA pull-down eluates using anti-Myc antibodies demonstrates a higher binding specificity of NONO-WT for CAGGCAGG RNA repeats compared to the S147A mutant in HEK293T cells. E Quantification of the SETMAR-L/SETMAR-S ratio in HEK293T cells by qPCR. F Left panel: RNA FISH analysis shows that O-GlcNAcylation of NONO promotes the expression of SETMAR-L. RNA FISH was conducted to visualize SETMAR-L (green) and SETMAR-S (red) in NONO-knockdown HEK293T cells stably reconstituted with either WT NONO or the S147A mutant. DAPI staining of the nuclei is shown in blue. Scale bar, 20 μm. Right panel: Quantification of fluorescence intensity from FISH. Data from three independent experiments are presented as mean ± SD. Scale bar, 20 μm

Techniques: Alternative Splicing, Transfection, Mutagenesis, Immunoprecipitation, Western Blot, Plasmid Preparation, Amplification, Negative Control, Binding Assay, Expressing, Knockdown, Stable Transfection, Staining, Fluorescence

Journal: Genome Biology

Article Title: O-GlcNAcylation of NONO mediates alternative splicing of SETMAR and facilitates NHEJ repair

doi: 10.1186/s13059-026-03930-5

Figure Lengend Snippet: O-GlcNAcylation of NONO facilitates H3K36me2 enrichment at DSBs and recruits Ku70 for NHEJ. A Left panel: O-GlcNAcylation at Ser147 stabilizes Ku70 recruitment at DNA damage sites. U2OS cells were transfected with GFP-Ku70, along with an empty vector, SFB-NONO WT, or the S147A mutant. Representative images demonstrated the dynamic recruitment of GFP-Ku70 to DNA damage sites. Data were presented as mean ± SEM. Scale bar, 10 μm. Right panel: Quantification of the time course of GFP-Ku70 recruitment following laser microirradiation. B Upper panel: O-GlcNAcylation at Ser147 stabilizes the binding of Ku70 to chromatin. HEK293T cells were transfected with an empty vector, SFB-NONO WT, or the S147A mutant and exposed to 10 Gy of IR. Following IR, cells were allowed to recover for different durations. Whole cell extract (WCE) and chromatin fractions were prepared and subjected to IP and Western blot analysis with the indicated antibodies. Lower panel: Quantification of Ku70 level, normalized to H3 levels in chromatin and presented as fold change relative to control samples ( n = 3 per condition). C Upper panel: Impact of O-GlcNAcylation at Ser147 on H3K36 demethylation levels. Stable HEK293T cells with NONO knockdown were reconstituted with shRNA-resistant NONO WT or S147A mutant. These cells were treated with IR and examined via western blot analysis with the indicated antibodies to assess H3K36me2 levels. Lower panel: Quantification of H3K36me2 level, normalized to H3 levels in chromatin and presented as fold change relative to control samples ( n = 3 per condition)

Techniques: Transfection, Plasmid Preparation, Mutagenesis, Binding Assay, Western Blot, Control, Knockdown, shRNA

Journal: Genome Biology

Article Title: O-GlcNAcylation of NONO mediates alternative splicing of SETMAR and facilitates NHEJ repair

doi: 10.1186/s13059-026-03930-5

Figure Lengend Snippet: O-GlcNAcylation of NONO promotes radioresistance in hepatocellular carcinoma. A Analysis of NONO O-GlcNAcylation was conducted in human liver epithelial cells (THLE2) and hepatocellular carcinoma cells (HepG2, HCCLM9, Huh7) usingIP/Western blotting with indicated antibodies. B Generation of stable HCCLM9 cells with NONO knockdown. Western blot analysis was performed to assess the efficiency of NONO knockdown using two shRNA constructs(shNONO #1 and shNONO #2). GAPDH was used as a loading control. C Upper panel: O-GlcNAcylation at Ser147 promotes cell survival post-IR treatment. NONO-knockdown HCCLM9 cells were transfected with SFB-NONO WT or S147A mutant and subjected to clonogenic survival assays post-IR treatment. Cells were treated with the indicated doses of IR and further incubated for 7–10 days. Lower panel: Quantitative analysis of clonogenic survival assays. D Schematic diagram illustrating the radiotherapy process for NTG mice. Mice injected with control or NONO knockdown cells, as well as cells reconstituted with NONO WT or S147A and wereexposed to 8 Gy of IR twice. E O-GlcNAcylation of NONO enhances radioresistance. Mice were subcutaneously injected with 7 × 10 6 control or NONO knockdown cells, or cells reconstituted with NONO WT or S147A, and exposed to 8 Gy of IR twice or not, when tumors reached a similar size (about 100 mm. 3 ). Representative images of xenograft tumors are shown ( n = 6/group). F Quantification analysis of xenograft tumor volumes from ( E ). Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA. ** P < 0.01, *** P < 0.001. G Hematoxylin and eosin (H&E) staining and IHC analysis of NONO and H3K36me2 in xenograft tumors were performed, comparing controland NONO knockdown groups, as well as tumors reconstituted with NONO WT or S147A ( n = 6). Scale bar, 30 μm. H Quantification of H3K36me2 levels from ( G ). Data are presented as mean ± SD. Statistical significance was determined using one-way ANOVA. ** P < 0.01, *** P < 0.001

Techniques: Western Blot, Knockdown, shRNA, Construct, Control, Transfection, Mutagenesis, Incubation, Injection, Staining

Journal: Genome Biology

Article Title: O-GlcNAcylation of NONO mediates alternative splicing of SETMAR and facilitates NHEJ repair

doi: 10.1186/s13059-026-03930-5

Figure Lengend Snippet: Proposed working model of NONO O-GlcNAcylation regulating NHEJ-mediated DNA damage repair

Techniques:

(A) Schematic illustration of experimental flows for proteomics analysis of NSD1-PWWP2’s interactomes. (B) Unique proteins detected by LC-MS and plotted by peptide-spectrum match (PSM) scores against percentage of coverage using DIPG13 (top) and HEK293T (bottom) cells. (C) Illustration of annotated functional domains of NONO. (D) GST pulldown assay of HA-tagged NONO using NSD1-PWWP2 as the bait. Left, pulldown of HA-tagged N-NONO or C-NONO using GST alone or GST-NSD1-PWWP2 followed by western blot of GST and HA. Right, pulldown of HA-tagged N-NONO using GST-NSD1-PWWP2 or GST-NSD1-PWWP2–4A mutant followed by western blot of GST and HA.

Journal: Cell reports

Article Title: Paraspeckle protein NONO regulates active chromatin by allosterically stimulating NSD1

doi: 10.1016/j.celrep.2025.116247

Figure Lengend Snippet: (A) Schematic illustration of experimental flows for proteomics analysis of NSD1-PWWP2’s interactomes. (B) Unique proteins detected by LC-MS and plotted by peptide-spectrum match (PSM) scores against percentage of coverage using DIPG13 (top) and HEK293T (bottom) cells. (C) Illustration of annotated functional domains of NONO. (D) GST pulldown assay of HA-tagged NONO using NSD1-PWWP2 as the bait. Left, pulldown of HA-tagged N-NONO or C-NONO using GST alone or GST-NSD1-PWWP2 followed by western blot of GST and HA. Right, pulldown of HA-tagged N-NONO using GST-NSD1-PWWP2 or GST-NSD1-PWWP2–4A mutant followed by western blot of GST and HA.

Article Snippet: NONO rabbit polyclonal , ProteinTech , Cat# 11058-1-AP.

Techniques: Liquid Chromatography with Mass Spectroscopy, Functional Assay, GST Pulldown Assay, Western Blot, Mutagenesis

(A) Demonstration of recombinant protein expression and purification, including NSD1, NSD1 PWWP2–4A , N-NONO, and recombinant di-nucleosomes by Coomassie blue staining. (B) HMT assays of full-length NSD1 or NSD1 PWWP2–4A mutant in an incremental titration of 62.5, 125, and 250 nM. Top: quantifications of autoradiographic signals normalized to NSD1 alone (the second lane). Middle: representative autoradiographic images for stably incorporated [ 3 H]. Bottom: Coomassie blue staining of total nucleosomes. Data are presented as mean ± SEM. (C) HMT assays of 60 nM full-length NSD1 with an incremental titration of N-NONO at 0, 240, 530, 880, and 1760 nM. Top: quantifications of autoradiographic signals normalized to NSD1 alone (the second lane). Middle: representative autoradiographic images for stably incorporated [ 3 H]. Bottom: Coomassie blue staining of total nucleosomes. Data are presented as mean ± SEM. (D) HMT assays of 0.25 μM NSD1 PWWP2–4A mutant with an incremental titration of N-NONO at 0, 530, 880, and 1760 nM. Top: quantifications of autoradiographic signals normalized to NSD1 alone (the second lane). Middle: representative autoradiographic images for stably incorporated [ 3 H]. Bottom, Coomassie blue staining of total nucleosomes. Data are presented as mean ± SEM.

Journal: Cell reports

Article Title: Paraspeckle protein NONO regulates active chromatin by allosterically stimulating NSD1

doi: 10.1016/j.celrep.2025.116247

Figure Lengend Snippet: (A) Demonstration of recombinant protein expression and purification, including NSD1, NSD1 PWWP2–4A , N-NONO, and recombinant di-nucleosomes by Coomassie blue staining. (B) HMT assays of full-length NSD1 or NSD1 PWWP2–4A mutant in an incremental titration of 62.5, 125, and 250 nM. Top: quantifications of autoradiographic signals normalized to NSD1 alone (the second lane). Middle: representative autoradiographic images for stably incorporated [ 3 H]. Bottom: Coomassie blue staining of total nucleosomes. Data are presented as mean ± SEM. (C) HMT assays of 60 nM full-length NSD1 with an incremental titration of N-NONO at 0, 240, 530, 880, and 1760 nM. Top: quantifications of autoradiographic signals normalized to NSD1 alone (the second lane). Middle: representative autoradiographic images for stably incorporated [ 3 H]. Bottom: Coomassie blue staining of total nucleosomes. Data are presented as mean ± SEM. (D) HMT assays of 0.25 μM NSD1 PWWP2–4A mutant with an incremental titration of N-NONO at 0, 530, 880, and 1760 nM. Top: quantifications of autoradiographic signals normalized to NSD1 alone (the second lane). Middle: representative autoradiographic images for stably incorporated [ 3 H]. Bottom, Coomassie blue staining of total nucleosomes. Data are presented as mean ± SEM.

Article Snippet: NONO rabbit polyclonal , ProteinTech , Cat# 11058-1-AP.

Techniques: Recombinant, Expressing, Purification, Staining, Mutagenesis, Titration, Stable Transfection

(A) Overlay of meta-analysis profiling of H3K36me2 ChIP-seq signals at all genes within a window of −10 kb of TSS to +10 kb of TES in WT and NONO-KO E14-mESC. Representative track images are shown at the bottom. (B) Individual meta-analysis profiling and heatmaps of H3K36me2 ChIP-seq in WT and NONO-KO mESCs. Left: ChIP-seq signals from WT cells were presented at all genes within a −10 kb of TSS to +10 kb of TES window, and NONO-KO cells were aligned to WT cells. Right: ChIP-seq signals were ranked by max peak value and aligned to the centers. (C) qPCR quantification of NEAT1 RNA expression levels in WT and NEAT1 CRISPRi cells. Signals were normalized by GAPDH . n = 5 for each condition. p value was calculated by Student’s t test. Data are presented as mean ± SEM. (D) Immunofluorescence staining of NONO in WT and NEAT1 CRISPRi HEK293T cells. Images were captured under a 63× objective, and the puncta of nuclear paraspeckles were highlighted by red triangles. Scale bars, 50 μm. (E) Quantifications of (D). Nuclear paraspeckles are present in individual WT ( n = 24) and NEAT1 CRISPRi ( n = 40) HEK293T cells. The p value is calculated by chi-squared test. (F) Overlay of meta-analysis profiling of H3K36me2 ChIP-seq signals at all genes within a window of −10 kb of TSS to +10 kb of TES in WT and NEAT1 CRISPRi HEK293T cells. Representative track images are shown at the bottom. (G) Individual meta-analysis profiling and heatmaps of H3K36me2 ChIP-seq in WT and NEAT1 CRISPRi HEK293T cells. ChIP-seq signals from WT cells were presented at all genes within a −10 kb of TSS to +10 kb of TES window, and NONO-KO cells were aligned to WT cells. Right: ChIP-seq signals were ranked by max peak value and aligned to the centers.

Journal: Cell reports

Article Title: Paraspeckle protein NONO regulates active chromatin by allosterically stimulating NSD1

doi: 10.1016/j.celrep.2025.116247

Figure Lengend Snippet: (A) Overlay of meta-analysis profiling of H3K36me2 ChIP-seq signals at all genes within a window of −10 kb of TSS to +10 kb of TES in WT and NONO-KO E14-mESC. Representative track images are shown at the bottom. (B) Individual meta-analysis profiling and heatmaps of H3K36me2 ChIP-seq in WT and NONO-KO mESCs. Left: ChIP-seq signals from WT cells were presented at all genes within a −10 kb of TSS to +10 kb of TES window, and NONO-KO cells were aligned to WT cells. Right: ChIP-seq signals were ranked by max peak value and aligned to the centers. (C) qPCR quantification of NEAT1 RNA expression levels in WT and NEAT1 CRISPRi cells. Signals were normalized by GAPDH . n = 5 for each condition. p value was calculated by Student’s t test. Data are presented as mean ± SEM. (D) Immunofluorescence staining of NONO in WT and NEAT1 CRISPRi HEK293T cells. Images were captured under a 63× objective, and the puncta of nuclear paraspeckles were highlighted by red triangles. Scale bars, 50 μm. (E) Quantifications of (D). Nuclear paraspeckles are present in individual WT ( n = 24) and NEAT1 CRISPRi ( n = 40) HEK293T cells. The p value is calculated by chi-squared test. (F) Overlay of meta-analysis profiling of H3K36me2 ChIP-seq signals at all genes within a window of −10 kb of TSS to +10 kb of TES in WT and NEAT1 CRISPRi HEK293T cells. Representative track images are shown at the bottom. (G) Individual meta-analysis profiling and heatmaps of H3K36me2 ChIP-seq in WT and NEAT1 CRISPRi HEK293T cells. ChIP-seq signals from WT cells were presented at all genes within a −10 kb of TSS to +10 kb of TES window, and NONO-KO cells were aligned to WT cells. Right: ChIP-seq signals were ranked by max peak value and aligned to the centers.

Article Snippet: NONO rabbit polyclonal , ProteinTech , Cat# 11058-1-AP.

Techniques: ChIP-sequencing, RNA Expression, Immunofluorescence, Staining

(A) Meta-analysis profiling and heatmaps of NSD1 ChIP-seq signals at all genes within a window of −10 kb of TSS to +10 kb of TES in WT and NONO-KO HEK293T cells. NONO-KO is aligned to WT. (B) Meta-analysis profiling and heatmaps of NONO ChIP-seq signals at all genes within a window of −10 kb of TSS to +10 kb of TES in WT and NSD1-KO HEK293T cells. NSD1-KO is aligned to WT. (C) Meta-analysis profiling and heatmaps of NSD1 ChIP-seq signals at all genes within a window of −10 kb of TSS to +10 kb of TES in WT and NONO-KO mESC cells. NONO-KO is aligned to WT. (D) Meta-analysis profiling and heatmaps of NONO ChIP-seq signals at all genes within a window of −10 kb of TSS to +10 kb of TES in WT and NSD1-KO mESC cells. NSD1-KO is aligned to WT.

Journal: Cell reports

Article Title: Paraspeckle protein NONO regulates active chromatin by allosterically stimulating NSD1

doi: 10.1016/j.celrep.2025.116247

Figure Lengend Snippet: (A) Meta-analysis profiling and heatmaps of NSD1 ChIP-seq signals at all genes within a window of −10 kb of TSS to +10 kb of TES in WT and NONO-KO HEK293T cells. NONO-KO is aligned to WT. (B) Meta-analysis profiling and heatmaps of NONO ChIP-seq signals at all genes within a window of −10 kb of TSS to +10 kb of TES in WT and NSD1-KO HEK293T cells. NSD1-KO is aligned to WT. (C) Meta-analysis profiling and heatmaps of NSD1 ChIP-seq signals at all genes within a window of −10 kb of TSS to +10 kb of TES in WT and NONO-KO mESC cells. NONO-KO is aligned to WT. (D) Meta-analysis profiling and heatmaps of NONO ChIP-seq signals at all genes within a window of −10 kb of TSS to +10 kb of TES in WT and NSD1-KO mESC cells. NSD1-KO is aligned to WT.

Article Snippet: NONO rabbit polyclonal , ProteinTech , Cat# 11058-1-AP.

Techniques: ChIP-sequencing

(A) Neural progenitor cell (NPC) differentiation of WT, NSD1-KO, and NONO-KO E14-mESCs. Top: representative images of embryoid bodies (EBs) undergoing NPC differentiation after 3 days of retinoic acid (RA) treatment. Bottom, quantifications of fully differentiated, partially differentiated, or non-differentiated EBs. Scale bars, 500 μm. (B) Heatmaps of differential gene expression analysis in WT, NSD1-KO, and NONO-KO cells treated with RA for 0, 3, or 6 days using RNA-seq. A total of 252 genes associated with neural development and 102 genes associated with stem cell differentiation were presented. (C) Heatmaps of significant changes of gene set enrichment analysis signatures, including stem cell differentiation and neural lineage gene sets in WT compared to NSD1-KO and NONO-KO E14-mESC cells undergoing RA-induced NPC differentiation. (D) Boxplots of log2 fold changes in gene expression using the experimental conditions shown in (B). The box and whisker represent 95%, the third quartile, the median, the first quartile, and 5% distribution of genes. Data are presented as mean ± SEM. p values were calculated by Wilcoxon test ** p < 0.01; *** p < 0.001; and **** p < 0.0001.

Journal: Cell reports

Article Title: Paraspeckle protein NONO regulates active chromatin by allosterically stimulating NSD1

doi: 10.1016/j.celrep.2025.116247

Figure Lengend Snippet: (A) Neural progenitor cell (NPC) differentiation of WT, NSD1-KO, and NONO-KO E14-mESCs. Top: representative images of embryoid bodies (EBs) undergoing NPC differentiation after 3 days of retinoic acid (RA) treatment. Bottom, quantifications of fully differentiated, partially differentiated, or non-differentiated EBs. Scale bars, 500 μm. (B) Heatmaps of differential gene expression analysis in WT, NSD1-KO, and NONO-KO cells treated with RA for 0, 3, or 6 days using RNA-seq. A total of 252 genes associated with neural development and 102 genes associated with stem cell differentiation were presented. (C) Heatmaps of significant changes of gene set enrichment analysis signatures, including stem cell differentiation and neural lineage gene sets in WT compared to NSD1-KO and NONO-KO E14-mESC cells undergoing RA-induced NPC differentiation. (D) Boxplots of log2 fold changes in gene expression using the experimental conditions shown in (B). The box and whisker represent 95%, the third quartile, the median, the first quartile, and 5% distribution of genes. Data are presented as mean ± SEM. p values were calculated by Wilcoxon test ** p < 0.01; *** p < 0.001; and **** p < 0.0001.

Article Snippet: NONO rabbit polyclonal , ProteinTech , Cat# 11058-1-AP.

Techniques: Gene Expression, RNA Sequencing, Cell Differentiation, Whisker Assay

Journal: iScience

Article Title: C9orf72 poly-PR forms anisotropic condensates causative of nuclear TDP-43 pathology

doi: 10.1016/j.isci.2024.110937

Figure Lengend Snippet:

Article Snippet: NONO (rabbit polyclonal) , Proteintech , Cat# 11058-1-AP; RRID: AB_2152167.

Techniques: Labeling, Virus, Recombinant, Transfection, Reverse Transcription, Lysis, Western Blot, Staining, Derivative Assay, Plasmid Preparation, Software, Imaging, Microscopy, Real-time Polymerase Chain Reaction

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