Journal: bioRxiv

Article Title: Age-dependent loss of cohesion protection in human oocytes

doi: 10.1101/2023.01.13.523952

Figure Lengend Snippet: (A) Sgo2 localization in GV, GVBD, metaphase I, telophase I, and metaphase II stage oocytes. Oocytes were immunostained with antibodies against Sgo2 (green), CenpC (inner kinetochores, magenta), a-tubulin (microtubules, orange) and counter stained with Hoechst (blue). White circles with dashed lines represent the oocyte circumference. White boxes with dashed lines represent chromosomes masses that have been further magnified below. (B) Representative line scans showing distinction between bridge and centromeric Sgo2 pools. A line was drawn as shown on individual sister chromatid pairs from metaphase II oocytes stained as in (A) and the fluorescence profile of Sgo2 is shown in the graph. (C) Comparison of Sgo2 localization at the pericentromeric bridge in metaphase II oocytes from women stratified by age (≤35 years or >35 years) undergoing ICSI or IVF treatment. Plots show median (dashed black line), 25th and 75th percentiles (dotted black lines). n.s. Not significant (Kruskal-Wallis test).

Article Snippet: Spreads were incubated in mouse monoclonal anti-PP2A Catalytic α antibody (1:50; BD Biosciences, 610556), rabbit polyclonal anti-SGOL2 antibody (1:200; Novus Biologicals, NBP1-83567), mouse polyclonal anti Rec8 antibody (1:50; Novus Biologicals, H00009985-B01P) human anti-CREST antibody/ antisera (1:50 Antibodies Incorporated, 15-235) and guinea pig polyclonal antibody anti-CenpC antibody (1:200; MBL, PB030) diluted in 3% BSA, 7% FBS in PBST.

Techniques: Staining, Fluorescence, Comparison

Journal: bioRxiv

Article Title: Age-dependent loss of cohesion protection in human oocytes

doi: 10.1101/2023.01.13.523952

Figure Lengend Snippet: (A-D) Human Sgo2 localizes to centromeric cups and an inter-sister pericentromere bridge in metaphase I oocytes and metaphase II oocytes of younger women, but is lost from the bridge with age. Representative images of human metaphase I (A and B) and metaphase II oocytes (C and D) from younger (age 30 and 29 years) and older women (age 40 and 39 years). Sgo2 (green), inner kinetochores (CenpC, magenta), microtubules (a-tubulin, orange) and chromosomes (Hoeschst, blue) are shown. White dashed line boxes indicate chromosomes further magnified in (B) metaphase I and (D) metaphase II. Centromeric localization of Sgo2 is bounded by dashed lines. Interpretations of localizations observed are indicated in the schematics below. (E and F) The percentage of chromatids per oocyte with Sgo2 localization at the pericentromeric bridge was scored relative to the woman’s age at MI (E) and MII (F). Data were fit to a Sigmodal, 4PL curve (metaphase I; R 2 = 0.84, metaphase II; R 2 = 0.67). Oocytes shown in (A-D) are labelled (green circles). (G) Centromeric Sgo2 signal is not significantly altered with age. The relative intensity of the centromeric pool of Sgo2 in metaphase II oocytes from younger (age ≤ 35 years) or older women (age >35 years) was measured in arbitrary units (A.U.) relative to the inner kinetochore (CENP-C) or centromere (CREST) markers. Plots show median (dashed black line), 25th and 75th percentiles (dotted black lines). P values were calculated using the Mann-Whitney test: CenpC ( P = 0.37) and CREST ( P = 0.15). n.s. = not significant. (H) Schematic summarises Sgo2 localization in younger women in metaphase I and II oocytes.

Article Snippet: Spreads were incubated in mouse monoclonal anti-PP2A Catalytic α antibody (1:50; BD Biosciences, 610556), rabbit polyclonal anti-SGOL2 antibody (1:200; Novus Biologicals, NBP1-83567), mouse polyclonal anti Rec8 antibody (1:50; Novus Biologicals, H00009985-B01P) human anti-CREST antibody/ antisera (1:50 Antibodies Incorporated, 15-235) and guinea pig polyclonal antibody anti-CenpC antibody (1:200; MBL, PB030) diluted in 3% BSA, 7% FBS in PBST.

Techniques: MANN-WHITNEY

Journal: bioRxiv

Article Title: Age-dependent loss of cohesion protection in human oocytes

doi: 10.1101/2023.01.13.523952

Figure Lengend Snippet: (A-F) Loss of Sgo2 at the pericentromeric bridge is associated with increased inter-sister kinetochore distance in metaphase I and metaphase II. (A) Inter-sister kinetochore distance (white dashed arrows) was determined on metaphase I chromosomes and related to the presence of the Sgo2 bridge. (B) Increase in inter-sister kinetochore distance at metaphase I with female age for chromosomes with a Sgo2 bridge (blue) or no bridge (green). Data was fit to a linear regression (R 2 = 0.81; P <0.0001). (C) Inter-sister kinetochore distance for sister chromatid pairs with a Sgo2 bridge or no bridge at metaphase I in younger (age ≤ 35 years) or older women (age >35 years). Plots show median (dashed black line), 25th and 75th percentiles (dotted black lines). Statistical analyses were performed using the Kruskal-Wallis test (**** P <0.0001). (D) Inter-sister kinetochore distance (white dashed arrows) was determined on metaphase II chromosomes and related to the presence of the Sgo2 bridge. (E) Increase in inter-sister kinetochore distance at metaphase II with female age for sister chromatid pairs with a Sgo2 bridge (blue) or no bridge (green). Data was fit to a linear regression (R 2 = 0.47; P <0.0001). (F) Inter-sister kinetochore distance where the Sgo2 bridge is present or absent at metaphase II in younger (age ≤ 35 years) or older women (age >35 years). Statistical analyses were performed using the Kruskal-Wallis test (**** P <0.0001, ** P = 0.0017). Plots show median (dashed black line), 25th and 75th percentiles (dotted black lines). (G) Centromeric Sgo2 does not correlate with inter-sister kinetochore distance. The relative intensity of the centromeric pool of Sgo2 metaphase II oocytes was measured in arbitrary units (A.U.) relative to the kinetochore markers CenpC ( P =0.23; R 2 = 0.0019) and CREST ( P = 0.052; R 2 = 0.012). Data was fit to a linear regression.

Article Snippet: Spreads were incubated in mouse monoclonal anti-PP2A Catalytic α antibody (1:50; BD Biosciences, 610556), rabbit polyclonal anti-SGOL2 antibody (1:200; Novus Biologicals, NBP1-83567), mouse polyclonal anti Rec8 antibody (1:50; Novus Biologicals, H00009985-B01P) human anti-CREST antibody/ antisera (1:50 Antibodies Incorporated, 15-235) and guinea pig polyclonal antibody anti-CenpC antibody (1:200; MBL, PB030) diluted in 3% BSA, 7% FBS in PBST.

Techniques:

Journal: bioRxiv

Article Title: Age-dependent loss of cohesion protection in human oocytes

doi: 10.1101/2023.01.13.523952

Figure Lengend Snippet: (A) Representative image showing presence of single chromatids (arrows) in human metaphase II oocytes from an older (age 40) women, compared to a younger (age 30) woman without single chromatids. White boxes with dashed lines indicate examples of paired and single chromosomes in oocyte 2. Sgo2 (green), inner kinetochores (CenpC; magenta), microtubules (a-tubulin; orange) and chromosomes (Hoechst; blue) are shown. (B) Increase of single chromatids in metaphase II oocytes with maternal age. The number of single chromatids was scored relative to woman’s age. Data were fit to a Sigmodal, 4PL curve (R 2 = 0.57). Oocytes used in representative images are labelled in the graphs. (C) Increased number of single chromatids in metaphase II oocytes with an increased fraction of sister chromatid pairs lacking a Sgo2 bridge. Data were fit to a Sigmodal, 4PL curve (R 2 = 0.60). (D) The relative intensity of the centromeric pool of Sgo2 between paired and single chromatids was measured only from oocytes that had single chromatids. Sgo2 intensity was measured in arbitrary units (A.U.) relative to the kinetochore markers CenpC ( P = 0.96, Mann-Whitney test) and CREST ( P = 0.24, Mann-Whitney test). Plots show median (dashed black line), 25th and 75th percentiles (dotted black lines). P values were calculated using the Mann-Whitney test. n.s., Not significant.

Article Snippet: Spreads were incubated in mouse monoclonal anti-PP2A Catalytic α antibody (1:50; BD Biosciences, 610556), rabbit polyclonal anti-SGOL2 antibody (1:200; Novus Biologicals, NBP1-83567), mouse polyclonal anti Rec8 antibody (1:50; Novus Biologicals, H00009985-B01P) human anti-CREST antibody/ antisera (1:50 Antibodies Incorporated, 15-235) and guinea pig polyclonal antibody anti-CenpC antibody (1:200; MBL, PB030) diluted in 3% BSA, 7% FBS in PBST.

Techniques: MANN-WHITNEY

Journal: bioRxiv

Article Title: Age-dependent loss of cohesion protection in human oocytes

doi: 10.1101/2023.01.13.523952

Figure Lengend Snippet: (A and B) Chromosome spreads of metaphase II-arrested human oocytes were stained with antibodies against Sgo2 (green), inner kinetochores (CenpC, magenta) and cohesin (Rec8, orange). Chromosomes were stained with Hoechst (blue). (A) Representative images with white dashed line boxes indicating representative chromosome figures shown at higher magnification on the right. (B) Localization of Rec8 in the pericentromere bridge was scored for sister chromatid pairs with and without a Sgo2 bridge. Schematic representations of the data are shown below. (C and D) Chromosome spreads of metaphase II-arrested human oocytes were stained with antibodies against Sgo2 (green), inner kinetochores (CenpC, magenta) and PP2A (orange). Chromosomes were stained with Hoechst (blue). (C) Representative images with white dashed line boxes indicating representative chromosome figures shown at higher magnification on the right. (D) Localization of PP2A in the pericentromere bridge was scored for sister chromatid pairs with and without a Sgo2 bridge. Schematic representations of the data are shown below.

Article Snippet: Spreads were incubated in mouse monoclonal anti-PP2A Catalytic α antibody (1:50; BD Biosciences, 610556), rabbit polyclonal anti-SGOL2 antibody (1:200; Novus Biologicals, NBP1-83567), mouse polyclonal anti Rec8 antibody (1:50; Novus Biologicals, H00009985-B01P) human anti-CREST antibody/ antisera (1:50 Antibodies Incorporated, 15-235) and guinea pig polyclonal antibody anti-CenpC antibody (1:200; MBL, PB030) diluted in 3% BSA, 7% FBS in PBST.

Techniques: Staining

Journal: bioRxiv

Article Title: Age-dependent loss of cohesion protection in human oocytes

doi: 10.1101/2023.01.13.523952

Figure Lengend Snippet: (A-D) Inhibition of Mps1 in metaphase II oocytes impairs Sgo2 localization. (A) Scheme of the experiment. Metaphase II eggs from women aged ≤ 36 years were treated with 500nM Reversine (to inhibit Mps1) or DMSO (control) overnight and fixed. (B) Representative images of control and reversine-treated metaphase II oocytes after immunostaining with antibodies against Sgo2 (green), CenpC (inner kinetochores, magenta) and a-tubulin (microtubules, orange). White boxes with dashed lines indicate chromosomes that have been further magnified below. (C) The percentage of chromatids per oocyte with Sgo2 localization at the pericentromeric bridge was scored in control and reversine-treated metaphase II oocytes (** P =0.0084; Welch’s t test). (D) The relative intensity of the centromeric pool of Sgo2 in metaphase II oocytes relative to CenpC in control and reversine-treated oocytes (**** P <0.0001; Mann-Whitney test). (E and F) Mps1 activity is required for Bub1 localization to kinetochores. Metaphase II oocytes treated with reversine as in (A) were immunostained with antibodies against Bub1 (green), CenpC (inner kinetochore; magenta), and counter stained with Hoechst (blue) to visualise chromosomes. (E) Representative images of Bub1 localisation in control and reversine-treated metaphase II oocytes. White boxes with dashed lines represent chromosomes that have been further magnified below. White dashed circles show examples of area selections for Bub1 intensity measurements. (F) The relative intensity of the centromeric Bub1 in metaphase II oocytes from control and reversine-treated oocytes relative to CenpC (**** P <0.0001; Mann-Whitney test). Plots show median (dashed black line), 25th and 75th percentiles (dotted black lines).

Article Snippet: Spreads were incubated in mouse monoclonal anti-PP2A Catalytic α antibody (1:50; BD Biosciences, 610556), rabbit polyclonal anti-SGOL2 antibody (1:200; Novus Biologicals, NBP1-83567), mouse polyclonal anti Rec8 antibody (1:50; Novus Biologicals, H00009985-B01P) human anti-CREST antibody/ antisera (1:50 Antibodies Incorporated, 15-235) and guinea pig polyclonal antibody anti-CenpC antibody (1:200; MBL, PB030) diluted in 3% BSA, 7% FBS in PBST.

Techniques: Inhibition, Control, Immunostaining, MANN-WHITNEY, Activity Assay, Staining

Journal: bioRxiv

Article Title: Age-dependent loss of cohesion protection in human oocytes

doi: 10.1101/2023.01.13.523952

Figure Lengend Snippet: Loss of Sgo2 localization upon inhibition of Bub1 in metaphase II oocytes. (A) Scheme of the experiment. Metaphase II oocytes from women aged ≤ 36 years were treated with 10μM Bay-320 (to inhibit Bub1 ) or DMSO (control) overnight and fixed. (B) Representative images of control and Bay-320-treated metaphase II oocytes after immunostaining with antibodies against Sgo2 (green), CenpC (inner kinetochores, magenta) and a-tubulin (microtubules, orange). White boxes with dashed lines indicate chromosomes that have been further magnified below. (C) The percentage of chromatids per oocyte with Sgo2 localization at the pericentromeric bridge was scored in control and Bay-320-treated metaphase II oocytes (**** P <0.0001, Mann-Whitney test). (D) The relative intensity of the centromeric pool of Sgo2 in metaphase II oocytes from control and Bay-320-treated oocytes were measured relative to CenpC. (****P <0.0001, Mann-Whitney test).Plots show median (dashed black line), 25th and 75th percentiles (dotted black lines).

Article Snippet: Spreads were incubated in mouse monoclonal anti-PP2A Catalytic α antibody (1:50; BD Biosciences, 610556), rabbit polyclonal anti-SGOL2 antibody (1:200; Novus Biologicals, NBP1-83567), mouse polyclonal anti Rec8 antibody (1:50; Novus Biologicals, H00009985-B01P) human anti-CREST antibody/ antisera (1:50 Antibodies Incorporated, 15-235) and guinea pig polyclonal antibody anti-CenpC antibody (1:200; MBL, PB030) diluted in 3% BSA, 7% FBS in PBST.

Techniques: Inhibition, Control, Immunostaining, MANN-WHITNEY

Journal: bioRxiv

Article Title: Age-dependent loss of cohesion protection in human oocytes

doi: 10.1101/2023.01.13.523952

Figure Lengend Snippet: The impact of MPS inhibition on protection of sister chromatid cohesion. (A) Schematic of experiment. Oocytes from women undergoing ICSI aged ≤ 36 years provided at or prior to GVBD were treated with 500nM reversine or DMSO, allowed to mature to metaphase II for up 24 h and fixed. (B) Representative images of oocytes treated with reversine from GVBD stage alongside control oocytes after immunostaining with antibodies against Sgo2 (green), CenpC (inner kinetochore, magenta), a-tubulin (microtubules, orange) and counter staining with Hoechst (chromosomes, blue). White boxes with dashed lines represent chromosomes that have been further magnified below. (C) The percentage of chromatids per oocyte with Sgo2 localization at the pericentromeric bridge was scored (*** P = 0.0001; Mann-Whitney test). (D) The relative intensity of the centromeric pool of Sgo2 relative to CenpC. (**** P <0.0001; Mann-Whitney test). (E) The number of single chromatids observed in metaphase II oocytes after treatment with reversine from GBVD compared to controls (*** P <0.0003; Mann-Whitney test). (F) Centromeric Sgo2 was measured in metaphase II oocytes that had single chromatids after reversine treatment from GVBD stage onwards, and values for paired and single chromatids were compared. Sgo2 intensity was measured in arbitrary units relative to CenpC. Plots show median (dashed black line), 25th and 75th percentiles (dotted black lines) ( P =0.57; Mann-Whitney test). (G) Model for age-dependent loss of cohesin protection. In oocytes from younger women, cohesion is maintained between sister chromatids. Mps1 at the kinetochore recruits Bub1 which phosphorylates histone H2A Thr120 in the centromeric chromatin. This phosphorylation allows for the recruitment of Sgo2 at the centromere and the pericentromeric bridge coincident with cohesin. During anaphase I, Sgo2 protects cohesin within the pericentromeric bridge from separase activity to ensures that sister chromatids remain together at metaphase II and disjoin accurately only upon fertilisation-triggered anaphase II. In oocytes from older women, pericentromeric cohesin deteriorates. This both increases inter-sister kinetochore distances and results in loss of Sgo2 from the pericentromeric bridge. In the absence of Sgo2 at the bridge, residual pericentromeric cohesin is vulnerable to separase-dependent cleavage in anaphase I. Consequently, sister chromatids risk premature separation and such single chromatids at metaphase II which will disjoin randomly at fertilisation resulting in increased incidences of aneuploid conceptions in older women.

Article Snippet: Spreads were incubated in mouse monoclonal anti-PP2A Catalytic α antibody (1:50; BD Biosciences, 610556), rabbit polyclonal anti-SGOL2 antibody (1:200; Novus Biologicals, NBP1-83567), mouse polyclonal anti Rec8 antibody (1:50; Novus Biologicals, H00009985-B01P) human anti-CREST antibody/ antisera (1:50 Antibodies Incorporated, 15-235) and guinea pig polyclonal antibody anti-CenpC antibody (1:200; MBL, PB030) diluted in 3% BSA, 7% FBS in PBST.

Techniques: Inhibition, Control, Immunostaining, Staining, MANN-WHITNEY, Phospho-proteomics, Activity Assay

Journal: eLife

Article Title: Division of labour between PP2A-B56 isoforms at the centromere and kinetochore

doi: 10.7554/eLife.42619

Figure Lengend Snippet:

Article Snippet: Antibody , Rabbit polyclonal anti-Sgo2 , Bethyl , Bethyl: A301-262A, RRID: AB_890650 , 1:1000.

Techniques: Recombinant, Clone Assay, Mutagenesis, Plasmid Preparation, Sequencing, Knock-In, Modification, Magnetic Beads, Software, Western Blot, Microscopy

Journal: Journal of Cancer

Article Title: High SGO2 predicted poor prognosis and high therapeutic value of lung adenocarcinoma and promoted cell proliferation, migration, invasion, and epithelial-to-mesenchymal transformation

doi: 10.7150/jca.86285

Figure Lengend Snippet: SGO2 increases the risk of poor prognosis for LUAD. (A, E, and H) Based on the TCGA database, SGO2 expression, ROC curve, and survival analysis in LUAD. (B, F, and I) Based on the GEO database, the validation dataset GSE30219 was used to verify the SGO2 expression, ROC curve, and survival analysis in LUAD. (C) The expression of SGO2 in LUAD and normal tissue. (D) The protein expression of SGO2 in LUAD and normal tissue. (G and J) Immunohistochemical staining of SGO2 in LUAD and normal tissue.

Article Snippet: The sections were incubated with a 1:1000 rabbit anti-human SGO2 antibody (Cat. # NBP1-83567; Novus, USA) overnight at a 4 °C in wet box.

Techniques: Expressing, Biomarker Discovery, Immunohistochemical staining, Staining

Journal: Journal of Cancer

Article Title: High SGO2 predicted poor prognosis and high therapeutic value of lung adenocarcinoma and promoted cell proliferation, migration, invasion, and epithelial-to-mesenchymal transformation

doi: 10.7150/jca.86285

Figure Lengend Snippet: High SGO2 is associated with a higher TNM stage of LUAD. (A) Based on the TCGA and GSE30219, the expression of SGO2 in T1 vs T2. (B) Based on the TCGA and GSE30219, the expression of SGO2 in N0 vs N1. (C) Based on the TCGA and GSE31210, the expression of SGO2 in stage I vs stage II + III. (D and G) Based on IHC, the protein expression and staining of SGO2 in T1 vs T2. (E and H) Based on IHC, the protein expression and staining of SGO2 in N0 vs N1. (F and I) Based on IHC, the protein expression and staining of SGO2 in stage I vs stage II + III.

Article Snippet: The sections were incubated with a 1:1000 rabbit anti-human SGO2 antibody (Cat. # NBP1-83567; Novus, USA) overnight at a 4 °C in wet box.

Techniques: Expressing, Staining

Journal: Journal of Cancer

Article Title: High SGO2 predicted poor prognosis and high therapeutic value of lung adenocarcinoma and promoted cell proliferation, migration, invasion, and epithelial-to-mesenchymal transformation

doi: 10.7150/jca.86285

Figure Lengend Snippet: SGO2 silencing inhibits the proliferation of lung cancer cells. (A)SGO2 expression after knockdown in A549 and H1299. (B)CCK8 assays were performed to detect A549 and H1299 proliferation. (C)Edu assays were performed to detect A549 and H1299 proliferation.

Article Snippet: The sections were incubated with a 1:1000 rabbit anti-human SGO2 antibody (Cat. # NBP1-83567; Novus, USA) overnight at a 4 °C in wet box.

Techniques: Expressing, Knockdown

Journal: Journal of Cancer

Article Title: High SGO2 predicted poor prognosis and high therapeutic value of lung adenocarcinoma and promoted cell proliferation, migration, invasion, and epithelial-to-mesenchymal transformation

doi: 10.7150/jca.86285

Figure Lengend Snippet: The downregulation of SGO2 affects migration, invasion, and EMT. (A) A Transwell assay was performed to examine the effect of SGO2 knockdown on A549 and H1299 cells. (C) A wound healing assay was performed to examine the effect of SGO2 knockdown on A549 and H1299 cells. (B and D) WB assay the expression levels of E-cadherin, N-cadherin, Vimentin, and Cytokeratin in A549 and H1299 to evaluate the effect on EMT after SGO2 knockdown.

Article Snippet: The sections were incubated with a 1:1000 rabbit anti-human SGO2 antibody (Cat. # NBP1-83567; Novus, USA) overnight at a 4 °C in wet box.

Techniques: Migration, Transwell Assay, Knockdown, Wound Healing Assay, Expressing

Journal: Journal of Cancer

Article Title: High SGO2 predicted poor prognosis and high therapeutic value of lung adenocarcinoma and promoted cell proliferation, migration, invasion, and epithelial-to-mesenchymal transformation

doi: 10.7150/jca.86285

Figure Lengend Snippet: SGO2 expression and tumor immune infiltration. (A) the expression of SGO2 was positively correlated with the infiltration of Memory B cells, Activated CD4+ memory T cells, and CD8+ T cells, while the infiltration of Memory B cells and Tregs decreased with increasing SGO2 expression. (B) Correlation of SGO2 expression with 28 distinct types of tumor-infiltrating immune cells based on ssGSEA.

Article Snippet: The sections were incubated with a 1:1000 rabbit anti-human SGO2 antibody (Cat. # NBP1-83567; Novus, USA) overnight at a 4 °C in wet box.

Techniques: Expressing

Journal: Journal of Cancer

Article Title: High SGO2 predicted poor prognosis and high therapeutic value of lung adenocarcinoma and promoted cell proliferation, migration, invasion, and epithelial-to-mesenchymal transformation

doi: 10.7150/jca.86285

Figure Lengend Snippet: High SGO2 has multiple therapeutic benefits. (A) Comparison of mutational landscapes of SGO between high cluster and low cluster, and Comparison of tumor mutation burden (TMB) between two clusters. (B) Comparison of first-line chemotherapy and targeted therapy drug targets of high and low SGO2 clusters. (C) Comparison of immunomodulatory drug targets of high and low SGO2 clusters.

Article Snippet: The sections were incubated with a 1:1000 rabbit anti-human SGO2 antibody (Cat. # NBP1-83567; Novus, USA) overnight at a 4 °C in wet box.

Techniques: Comparison, Mutagenesis

Journal: Scientific reports

Article Title: Shugosin 2 is a biomarker for pathological grading and survival prediction in patients with gliomas.

doi: 10.1038/s41598-021-97119-4

Figure Lengend Snippet: Figure 2. The expression level of SGO2 is related to the survival of patients with high-grade gliomas. The Kaplan–Meier survival curve analyzed from GEO profile (GDS1816/230165_at/SGO2) (a), TCGA (b), and CGGA (c). Data showed that patients with high expression of SGO2 had unfavorable survival outcome. (GDS1816/230165_at/SGO2, n = 77, p = 0.0011 by log-rank test, 95% CI 1.000–1.00, hazard ratio 1.001; TCGA, n = 343, p < 1 × 10–15 by log-rank test, 95% CI 0.87–5.30, hazard ratio 3.90; CGGA, n = 209, p = 1.23 × 10–11 by log- rank test, 95% CI 2.13–4.25, hazard ratio 3.01).

Article Snippet: The tissue microarrays were incubated with a polyclonal rabbit antihuman SGO2 antibody (HPA035163, Atlas Antibodies, Stockholm.

Techniques: Expressing

Journal: Scientific reports

Article Title: Shugosin 2 is a biomarker for pathological grading and survival prediction in patients with gliomas.

doi: 10.1038/s41598-021-97119-4

Figure Lengend Snippet: Figure 1. The expression level of SGO2 is related to pathological grading of gliomas. The SGO2 mRNA level in different grade of gliomas and non-tumor brain tissue from GEO profile (GDS1962/230165_at/SGO2) (a), TCGA (b), and CCGA (c). SGO2 expression was significantly higher in high-grade gliomas (Grade III and IV) than in low-grade gliomas (Grade II) and non-tumor control. The Y-axis indicates the SGO2 mRNA expression. The p value was adjusted by Bonferroni method in R software (version 3.0.1) between each group.

Article Snippet: The tissue microarrays were incubated with a polyclonal rabbit antihuman SGO2 antibody (HPA035163, Atlas Antibodies, Stockholm.

Techniques: Expressing, Control, Software

Journal: Scientific reports

Article Title: Shugosin 2 is a biomarker for pathological grading and survival prediction in patients with gliomas.

doi: 10.1038/s41598-021-97119-4

Figure Lengend Snippet: Figure 4. Validation of SGO2 protein expression in human gliomas and non-tumor brain tissue. Hematoxylin and eosin staining of non-tumor brain tissue (a), low grade (b) and high grade gliomas (c). The immunohistochemical staining of SGO2 on non-tumor brain tissue (d), low grade (e), and high grade gliomas (f) (scale bar: 50 μm). (g–i) The SGO2 immunostaining scores in normal brain tissue, low-grade glioma and high-grade glioma were statistically analyzed. The adjusted p value was calibrated between each group.

Article Snippet: The tissue microarrays were incubated with a polyclonal rabbit antihuman SGO2 antibody (HPA035163, Atlas Antibodies, Stockholm.

Techniques: Biomarker Discovery, Expressing, Staining, Immunohistochemical staining, Immunostaining

Journal: Scientific reports

Article Title: Shugosin 2 is a biomarker for pathological grading and survival prediction in patients with gliomas.

doi: 10.1038/s41598-021-97119-4

Figure Lengend Snippet: Figure 3. Validation of SGO2 mRNA and protein levels in glioma cell lines and normal brain tissue (a) qRT- PCR was performed to examine SGO2 mRNA expression and the quantitative results are shown in glioma cell lines. The relative expressions were normalized with normal brain. Bars mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.005 showed significant differences. Data are representative of three independent experiments. (b) Protein lysates of glioma cell lines, including U87MG, LN229, GBM8401, and U118MG were applied to SDS-PAGE and Western blot analysis to quantitate SGO2 protein expression (full length blot is presented in Supplementary Fig. 1). GAPDH served as a loading control.

Article Snippet: The tissue microarrays were incubated with a polyclonal rabbit antihuman SGO2 antibody (HPA035163, Atlas Antibodies, Stockholm.

Techniques: Biomarker Discovery, Quantitative RT-PCR, Expressing, SDS Page, Western Blot, Control

Journal: Scientific reports

Article Title: Shugosin 2 is a biomarker for pathological grading and survival prediction in patients with gliomas.

doi: 10.1038/s41598-021-97119-4

Figure Lengend Snippet: Figure 5. The effect of SGO2 on cell proliferation and apoptosis (a) The SGO2 knockdown model constructed by siRNA 25 nM transfection into LN229 and GBM8401 cell lines. The knockdown efficiency of SGO2 siRNA or control siRNA in infected LN229 and GBM8401 cells measured by RT-qPCR. Bars, mean ± SEM; *p < 0.05, **p < 0.01, ***p < 0.005 showed significant differences. Data are representative of three independent experiments. (b) LN229 and GBM8401 cells were transfected with 25 nM siRNA or siControl. Cell count was determined at the indicated time points. The data are expressed as the mean ± s.d.; n = 3; **p < 0.01, and ***p < 0.001. (c) LN229 and GBM8401 cell with siSGO2 or siControl transfection were labeled with BrdU then proceeded analysis by flow cytometry (**p < 0.01, ***p < 0.005). (d) Cell cycle analysis of LN229 and GBM8401 siSGO2 cells was determined by propidium iodide (PI) stain and flow cytometry. The data are expressed as the mean ± s.d.; n = 3; *p < 0.05, **p < 0.01. (e) Cell apoptosis analysis of LN 229 and GBM8401 siSGO2 cells were determined by tetraethylbenzimidazolylcarbocyanine iodide (JC-1) dye and flow cytometry.

Article Snippet: The tissue microarrays were incubated with a polyclonal rabbit antihuman SGO2 antibody (HPA035163, Atlas Antibodies, Stockholm.

Techniques: Knockdown, Construct, Transfection, Control, Infection, Quantitative RT-PCR, Cell Counting, Labeling, Flow Cytometry, Cell Cycle Assay, Staining

Journal: Scientific reports

Article Title: Shugosin 2 is a biomarker for pathological grading and survival prediction in patients with gliomas.

doi: 10.1038/s41598-021-97119-4

Figure Lengend Snippet: Figure 6. The effect of SGO2 knockdown on cell migration detected by wound-healing assays. Images and Quantitative analysis of LN229 (a) and GBM8401 (b) cells in the wound-healing assay. Data are presented as the mean ± SD (n = 3). *p < 0.05, **p < 0.01.

Article Snippet: The tissue microarrays were incubated with a polyclonal rabbit antihuman SGO2 antibody (HPA035163, Atlas Antibodies, Stockholm.

Techniques: Knockdown, Migration, Wound Healing Assay

Journal: Scientific reports

Article Title: Shugosin 2 is a biomarker for pathological grading and survival prediction in patients with gliomas.

doi: 10.1038/s41598-021-97119-4

Figure Lengend Snippet: Figure 7. The SGO2 protein–protein interaction (PPI) network. (a) In the PPI network established by STRING dataset, SGO2 is a hub protein. (b) The STRING dataset also predicted the association between SGO2, ARUKB, and FOXM1. (c) Protein lysates of LN229 and GBM8401 were applied to SDS-PAGE and Western blot to investigate the protein expression of AURKB and FOXM1(full length blot is presented in Supplementary Fig. 2). α-actinin served as a loading control.

Article Snippet: The tissue microarrays were incubated with a polyclonal rabbit antihuman SGO2 antibody (HPA035163, Atlas Antibodies, Stockholm.

Techniques: SDS Page, Western Blot, Expressing, Control