Journal: FEBS Open Bio

Article Title: Identification of nuclear phosphoproteins as novel tobacco markers in mouse lung tissue following short-term exposure to tobacco smoke

doi: 10.1016/j.fob.2014.08.002

Figure Lengend Snippet: Micrographs of the immunofluorescently stained lung tissues obtained from control and mice exposed to tobacco smoke for 7 days. Panel A shows micrographs of DAPI staining (left), OSF3 staining (center), and the merged image (right). Panel B shows micrographs of DAPI staining (left), SPTBN1 staining (center), and the merged image (right; magnification 400×).

Article Snippet: An anti-OSF3 antibody (AB41906; Abcam, Cambridge, UK) and an anti-SPTBN1 antibody (19722-1-AP, Protein Tech Group.

Techniques: Staining, Control

Journal: Nature Communications

Article Title: LMO7 deficiency reveals the significance of the cuticular plate for hearing function

doi: 10.1038/s41467-019-09074-4

Figure Lengend Snippet: LMO7 re-organizes the F-actin network. a Schematic illustration of functional domains in LMO7 and corresponding domain deletion constructs (ΔPDZ, ΔLIM, ΔCH and Δ1-353aa). b Confocal imaging analysis of LMO7-WT, ΔPDZ, ΔLIM, ΔCH and Δ1-353aa fused to GFP, overexpressed in COS-7 cells. ΔPDZ and ΔCH constructs re-organize F-actin in a manner similar to full-length LMO7. ΔLIM and Δ1-353aa constructs remain co-localized with F-actin, but lose F-actin condensing capacity. c Confocal imaging analysis of LMO7-GFP co-transfected with spectrin, actinin or NMII in COS-7 cells. Scale bars, 2.5 μm

Article Snippet: All Lmo7 deletion constructs were created by using Q5 Site-Directed Mutagenesis Kit (NEB). αII-Spectrin-MycDDk (RC226485) and βII-Spectrin-MycDDk (RC212868) were obtained from Origene.

Techniques: Functional Assay, Construct, Imaging, Transfection

Journal: Communications Biology

Article Title: IGF2BP3 recruits NUDT21 to regulate SPTBN1 alternative polyadenylation and drive ovarian cancer progression

doi: 10.1038/s42003-025-08097-6

Figure Lengend Snippet: A–D scRNA-seq analysis of two OC patients tissue samples and one normal ovarian tissue sample. A Composition of the three scRNA-seq datasets according to the UMAP plot. B UMAP plot of major cell types, with the cell number as well as the proportion of each type illustrated in ( C ). D Dot plot of key APA regulators in five major cell types. E Boxplot of key APA regulators in OC cells vs. fallopian tube epithelial cells. Data from the GEO database (GSE151316). F CPTAC database analysis of SPTBN1 protein expression (normal: n = 25, tumor: n = 100). G Validation of NUDT21 expression in clinical tissue samples (The upper panel represents 6 paired samples, while the lower panel represents 6 unpaired samples. F: Fallopian tube epithelial tissue, T: OC tissue). H Kaplan‒Meier curves of progression-free survival ( n = 614) and overall survival ( n = 655) in OC patients with high and low NUDT21 levels.

Article Snippet: The primary antibodies used were as follows: NUDT21 (Proteintech, 10322-1-AP), METTL3 (Proteintech, 15073-1-AP), IGF2BP3 (Proteintech, 14642-1-AP), CPSF6 (Proteintech, 15489-1-AP), TRIM21 (Proteintech, 12108-1-AP), EIF3C (Proteintech, 12733-1-AP), EIF3D (Proteintech, 10219-1-AP), U2AF2 (Proteintech, 15624-1-AP), SRSF3 (Proteintech, 10916-1-AP), SRSF6 (Proteintech, 11772-1-AP), FLAG-tag (MBL, PM020), SPTBN1 (Proteintech, 25681-1-AP), CDK1 (Proteintech, 19532-1-AP), CDK2 (Proteintech, 10122-1-AP), CDK4 (Proteintech, 11026-1-AP), CDK6 (Proteintech, 14052-1-AP), CDK7 (Proteintech, 27027-1-AP), CDK9 (Proteintech, 11705-1-AP), Cyclin B1 (Proteintech, 55004-1-AP), Cyclin A2 (Proteintech, 18202-1-AP) and GAPDH (Proteintech, 60004-1-Ig).

Techniques: Expressing, Biomarker Discovery

Journal: Communications Biology

Article Title: IGF2BP3 recruits NUDT21 to regulate SPTBN1 alternative polyadenylation and drive ovarian cancer progression

doi: 10.1038/s42003-025-08097-6

Figure Lengend Snippet: A Venn diagram of NUDT21 IP-MS data with or without RNaseA, the eukaryotic RBP database (EuRBPDB) and the RBP database for tumorigenesis (RBPTD). B Co-IP assay of NUDT21 and IGF2BP3 in A2780 OVCAR3 and SKOV3 cells. C–F PAS-seq analysis of NUDT21-knockdown OVCAR3 cells; density scatter plots of long and short genes ( C ); and boxplot of the proximal and distal PASs uses ( D ). KEGG enrichment analysis of long genes ( E ) and short genes ( F ). G and H Volcano plot of IGF2BP3-binding genes identified by RIP-seq ( G ) and KEGG enrichment analysis of these genes ( H ). I and J Volcano plot of NUDT21-binding genes identified by RIP-seq ( I ) and KEGG enrichment analysis of these genes ( J ). K Venn diagram of the candidate target genes screened via meRIP-seq, NUDT21 PAS-seq, NUDT21 RIP-seq, and IGF2BP3 RIP-seq data. L Information on four candidate target genes from 56 genes related to cancer and APA. M UCSC browser showing the long and short isoforms of SPTBN1 , in which the proximal polyA site (pPAS) is in the 32 (35 total) introns. N PAS-seq data for SPTBN1 genes with NUDT21 knockdown. The proximal and distal PASs were marked by colored boxes, and the locations are labeled at the bottom. O MeRIP-seq results showing enrichment of m6A sites on intron 32 of SPTBN1 .

Article Snippet: The primary antibodies used were as follows: NUDT21 (Proteintech, 10322-1-AP), METTL3 (Proteintech, 15073-1-AP), IGF2BP3 (Proteintech, 14642-1-AP), CPSF6 (Proteintech, 15489-1-AP), TRIM21 (Proteintech, 12108-1-AP), EIF3C (Proteintech, 12733-1-AP), EIF3D (Proteintech, 10219-1-AP), U2AF2 (Proteintech, 15624-1-AP), SRSF3 (Proteintech, 10916-1-AP), SRSF6 (Proteintech, 11772-1-AP), FLAG-tag (MBL, PM020), SPTBN1 (Proteintech, 25681-1-AP), CDK1 (Proteintech, 19532-1-AP), CDK2 (Proteintech, 10122-1-AP), CDK4 (Proteintech, 11026-1-AP), CDK6 (Proteintech, 14052-1-AP), CDK7 (Proteintech, 27027-1-AP), CDK9 (Proteintech, 11705-1-AP), Cyclin B1 (Proteintech, 55004-1-AP), Cyclin A2 (Proteintech, 18202-1-AP) and GAPDH (Proteintech, 60004-1-Ig).

Techniques: Protein-Protein interactions, Co-Immunoprecipitation Assay, Knockdown, Binding Assay, Labeling

Journal: Communications Biology

Article Title: IGF2BP3 recruits NUDT21 to regulate SPTBN1 alternative polyadenylation and drive ovarian cancer progression

doi: 10.1038/s42003-025-08097-6

Figure Lengend Snippet: A – C RT‒qPCR was used to detect the relative changes in the lengths of the target genes ICMT , ZEB1 , RNF213 and SPTBN1 in A2780, OVCAR3, and SKOV3 cells following NUDT21 ( A ), IGF2BP3 ( B ) and METTL3 ( C ) knockdown ( n = 4). D MeRIP‒qPCR of SPTBN1 m6A modification; RIP‒qPCR of SPTBN1 interaction with IGF2BP3 and NUDT21 ( n = 4). E and F RIP-qPCR was used to validate the relative fold change in SPTBN1 long and short isoform binding with NUDT21 in OVCAR3 cells (F) following IGF2BP3 knockdown ( E ) ( n = 4). G and H RIP-qPCR was used to validate the relative fold change in SPTBN1 long and short isoform binding with NUDT21 in OVCAR3 cells ( H ) following METTL3 knockdown ( G ) ( n = 3). I A dual-luciferase assay was performed on the 32nd intron of SPTBN1 with WT and m6A site mutations, which were both cotransfected with IGF2BP3 ( n = 4). J – L SPTBN1 long and short isoform protein levels were detected by Western blot in A2780, OVCAR3, and SKOV3 cells after NUDT21 ( J ), IGF2BP3 ( K ), and METTL3 ( L ) were knocked down. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The primary antibodies used were as follows: NUDT21 (Proteintech, 10322-1-AP), METTL3 (Proteintech, 15073-1-AP), IGF2BP3 (Proteintech, 14642-1-AP), CPSF6 (Proteintech, 15489-1-AP), TRIM21 (Proteintech, 12108-1-AP), EIF3C (Proteintech, 12733-1-AP), EIF3D (Proteintech, 10219-1-AP), U2AF2 (Proteintech, 15624-1-AP), SRSF3 (Proteintech, 10916-1-AP), SRSF6 (Proteintech, 11772-1-AP), FLAG-tag (MBL, PM020), SPTBN1 (Proteintech, 25681-1-AP), CDK1 (Proteintech, 19532-1-AP), CDK2 (Proteintech, 10122-1-AP), CDK4 (Proteintech, 11026-1-AP), CDK6 (Proteintech, 14052-1-AP), CDK7 (Proteintech, 27027-1-AP), CDK9 (Proteintech, 11705-1-AP), Cyclin B1 (Proteintech, 55004-1-AP), Cyclin A2 (Proteintech, 18202-1-AP) and GAPDH (Proteintech, 60004-1-Ig).

Techniques: Knockdown, Modification, Binding Assay, Luciferase, Western Blot

Journal: Communications Biology

Article Title: IGF2BP3 recruits NUDT21 to regulate SPTBN1 alternative polyadenylation and drive ovarian cancer progression

doi: 10.1038/s42003-025-08097-6

Figure Lengend Snippet: A Western blot analysis of total, long, and short isoform SPTBN1-knockdown cells. B–D CCK-8 ( B ) ( n = 4), colony formation ( C ) ( n = 3) and Transwell ( D ) ( n = 3) assays of A2780, OVCAR3, and SKOV3 cells infected with SPTBN1 total, long, or short isoform knockdown or negative control. E Gross xenograft model anatomy and tumor volume and tumor weight after subcutaneous injection of SPTBN1 total, long, and short isoform knockdown in OVCAR3 cells into nude mice ( n = 5). F Intraperitoneal tumor metastasis of SPTBN1 total, long, and short isoform-knockdown OVCAR3 cells injected into nude mice; the graphs show the number of metastatic nodules ( n = 5). G Western blot analysis of NUDT21-knockdown and SPTBN1 short isoform-overexpressing cells. H–J Overexpression of the SPTBN1 short isoform reversed the growth (H) (n = 4), proliferation ( I ) ( n = 3) and metastasis ( J ) ( n = 3) of OC cells in which NUDT21 was downregulated. *P < 0.05, **P < 0.01, ***P < 0.001.

Article Snippet: The primary antibodies used were as follows: NUDT21 (Proteintech, 10322-1-AP), METTL3 (Proteintech, 15073-1-AP), IGF2BP3 (Proteintech, 14642-1-AP), CPSF6 (Proteintech, 15489-1-AP), TRIM21 (Proteintech, 12108-1-AP), EIF3C (Proteintech, 12733-1-AP), EIF3D (Proteintech, 10219-1-AP), U2AF2 (Proteintech, 15624-1-AP), SRSF3 (Proteintech, 10916-1-AP), SRSF6 (Proteintech, 11772-1-AP), FLAG-tag (MBL, PM020), SPTBN1 (Proteintech, 25681-1-AP), CDK1 (Proteintech, 19532-1-AP), CDK2 (Proteintech, 10122-1-AP), CDK4 (Proteintech, 11026-1-AP), CDK6 (Proteintech, 14052-1-AP), CDK7 (Proteintech, 27027-1-AP), CDK9 (Proteintech, 11705-1-AP), Cyclin B1 (Proteintech, 55004-1-AP), Cyclin A2 (Proteintech, 18202-1-AP) and GAPDH (Proteintech, 60004-1-Ig).

Techniques: Western Blot, Knockdown, CCK-8 Assay, Infection, Negative Control, Injection, Over Expression

Journal: Communications Biology

Article Title: IGF2BP3 recruits NUDT21 to regulate SPTBN1 alternative polyadenylation and drive ovarian cancer progression

doi: 10.1038/s42003-025-08097-6

Figure Lengend Snippet: A Structural diagram of the SPTBN1 long and short isoforms. B Venn diagram of SPTBN1 IP-MS data for long and short isoforms. C and D KEGG enrichment analysis of short unique genes ( C ) and long unique genes ( D ). E Co-IP assay of SPTBN1 long and short isoform binding to CDKs with RNase. F Co-IP was used to detect the interaction between CDK1 and its complex (CDK2, CyclinA2/B1) after total and long isoform SPTBN1 knockdown (upper panel) or SPTBN1 overexpression (lower panel). G and H Cell cycle assays were performed with knockdown of total SPTBN1 and the SPTBN1 long isoform ( G ), NUDT21 ( H ), and the statistics were presented for G2/M ( n = 3). *P < 0.05, **P < 0.01, ***P < 0.001. I Diagram of the regulatory mechanism.

Article Snippet: The primary antibodies used were as follows: NUDT21 (Proteintech, 10322-1-AP), METTL3 (Proteintech, 15073-1-AP), IGF2BP3 (Proteintech, 14642-1-AP), CPSF6 (Proteintech, 15489-1-AP), TRIM21 (Proteintech, 12108-1-AP), EIF3C (Proteintech, 12733-1-AP), EIF3D (Proteintech, 10219-1-AP), U2AF2 (Proteintech, 15624-1-AP), SRSF3 (Proteintech, 10916-1-AP), SRSF6 (Proteintech, 11772-1-AP), FLAG-tag (MBL, PM020), SPTBN1 (Proteintech, 25681-1-AP), CDK1 (Proteintech, 19532-1-AP), CDK2 (Proteintech, 10122-1-AP), CDK4 (Proteintech, 11026-1-AP), CDK6 (Proteintech, 14052-1-AP), CDK7 (Proteintech, 27027-1-AP), CDK9 (Proteintech, 11705-1-AP), Cyclin B1 (Proteintech, 55004-1-AP), Cyclin A2 (Proteintech, 18202-1-AP) and GAPDH (Proteintech, 60004-1-Ig).

Techniques: Protein-Protein interactions, Co-Immunoprecipitation Assay, Binding Assay, Knockdown, Over Expression

Journal:

Article Title: Unilateral Blood Flow Decrease Induces Bilateral and Symmetric Responses in the Immature Brain

doi: 10.2353/ajpath.2009.090257

Figure Lengend Snippet: Unilateral transient CCA occlusion can induce casp3 cleavage but not its downstream subtracts. A: Representative Western blots showing cleaved casp3 fragment (17 kDa) both in the IL and CL 48 hours after uni-tCCAo + MCAo (model M1) and after uni-CCAo (model M2) as compared with sham-operated neocortical cytosolic extracts (n = 8 for each condition). Highly cleaved casp3 was present in the IL after model M1, but it also is increased in the CL cortex and after model M2 compared with sham (sh) cytosolic extracts (P < 0.005). The cytosolic marker β-actin was used as protein loading control. B: Upstream proapoptotic molecules are expressed after neonatal stroke (small units of casp2, casp8, and casp9). After uni-tCCAo, intermediate cleavage fragments of casp2 and cytosolic cytochrome c release were detected, as compared with sham animals. C, upper panel: Western blot showing calpain-cleaved fragments (150 to 145 kDa) and casp3-cleaved fragment (120 kDa) of α II-spectrin, and cleaved poly(ADP-ribose) polymerase-1 fragment (89 kDa). All these fragments were found in the IL extracts after model M1. In contrast, only the 150 kDa fragment was observed after model M2. C, lower panel: Quantification of the 150-kDa band (percentage between OD of the 150-kDa and total 240-kDa α II-spectrin). Data represent mean ± SEM (n = 6 each). *P < 0.05; **P < 0.005; and ***P < 0.0001.

Article Snippet: Antibodies against αII-spectrin (FG 6090; BIOMOL, Coger, Paris, France), poly(ADP-ribose) polymerase-1 (11835238001; Roche), and β-actin (clone AC-15; Sigma-Aldrich) were also used.

Techniques: Western Blot, Marker, Control