Journal: Journal of Heredity

Article Title: Differences in Cell Proliferation and Craniofacial Phenotype of Closely Related Species in the Pupfish Genus Cyprinodon

doi: 10.1093/jhered/esz074

Figure Lengend Snippet: Immunofluorescence light sheet imaging visualizes number and location of mitotic cells as indicated by assay for pH3 in the heads of hatching-age Cyprinodon. (A) Regions of the head analyzed for proliferating cells. (B) 3D reconstructions in lateral view of the head for the DAPI channel (gray, stains nuclei), the pH3 channel (red, dividing cells), and both channels merged. Note that pH3 positive cells (red dots) are mainly localized to ventral structures as shown in lateral view. (C, D) Single 2D images in (C) frontal view and (D) lateral view show pH3 cells (red dots) localizing to epithelial and mesenchyme tissues surrounding cartilage elements as exemplified by the ceratohyal. Cartilage cells are identifiable in the DAPI channel (gray) by their widely spaced nuclei indicative of large cuboidal cells. Cartilage elements can be identified by shape, and are outlined by the brightly labeled perichondrium, a dense layer of mesenchyme cells that surrounds cartilage elements. Inset shows region outlined by white box and arrows point to pH3 positive cells in perichondrium. Note also the clusters of pH3 positive cells in jaws (arrow head), especially around lateral edge of jaws as seen in panel C. Labels: bh, basihyal cartilage; br, brain; ch, ceratohyal cartilage; ey, eye; ga, gill arches; lj, lower jaw; pa, pharynx; pf, pectoral fin; uj, upper jaw.

Article Snippet: Cells were labeled for pH3 with primary rabbit anti-pH3 polyclonal antibody (MyBioSource, MBS2517807), and secondary antibody Alexa Fluor 568 goat anti-Rabbit (Thermo Fisher, A11011).

Techniques: Immunofluorescence, Imaging, Labeling

Journal: Journal of Heredity

Article Title: Differences in Cell Proliferation and Craniofacial Phenotype of Closely Related Species in the Pupfish Genus Cyprinodon

doi: 10.1093/jhered/esz074

Figure Lengend Snippet: Cell proliferation varies among species of Cyprinodon in different regions of the head at hatching. Shown are number of pH3 positive cells relative to either surface area of sampled tissue (A) or volume of sampled tissue (B) for the 3 regions of the head sampled plus the index of relative jaw proliferation (see text). Plotted are values for each sample and boxplots. Samples sizes are snail-eater N = 30, omnivore N = 28, scale-biter N = 19. Significance for all post hoc pairwise comparisons (Tukey) is shown above boxplots and corresponds to Supplementary Table S3. Note that levels of proliferation significantly vary between species when sampling the entire head (e.g., head region standardized to surface area) or regions of the head posterior to the jaws (post-jaw subset), but proliferation does not vary in the jaws (jaw subset). ns, not significant. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Cells were labeled for pH3 with primary rabbit anti-pH3 polyclonal antibody (MyBioSource, MBS2517807), and secondary antibody Alexa Fluor 568 goat anti-Rabbit (Thermo Fisher, A11011).

Techniques: Sampling

Journal: Oncology Letters

Article Title: Role of high ubiquitin-conjugating enzyme E2 expression as a prognostic factor in nasopharyngeal carcinoma

doi: 10.3892/ol.2022.13314

Figure Lengend Snippet: Increased expression levels of UBE2B in NPC tissues. Analysis of the public datasets (A) GSE12452 and (B) GSE68799 indicated increased expression levels of UBE2B in NPC compared to normal nasopharynx tissues. The heatmaps (left panel) present UBE2B transcription levels in each tissue sample and the scatter plots (right panel) indicate gene expression compared between normal nasopharyngeal mucosa and NPC tissues. The GSE12452 dataset contained mRNA signals from 10 non-NPC and 31 NPC tissues and the GSE68799 dataset contained 4 non-NPC and 42 NPC tissues. NPC, nasopharyngeal carcinoma; UBE2B, ubiquitin-conjugating enzyme E2 B.

Article Snippet: The slides were then washed with tris-buffered saline and incubated with a primary rabbit polyclonal antibody against UBE2B (1:100 dilution; GeneTex, Inc.) for 1 h. A ChemMate EnVision kit (DAKO; Agilent Technologies, Inc.) was applied to detect the primary antibody.

Techniques: Expressing, Gene Expression, Ubiquitin Proteomics

Journal: Oncology Letters

Article Title: Role of high ubiquitin-conjugating enzyme E2 expression as a prognostic factor in nasopharyngeal carcinoma

doi: 10.3892/ol.2022.13314

Figure Lengend Snippet: UBE2B has an important role in the carcinogenesis of NPC cells. (A) Western blot analysis demonstrated that UBE2B expression levels were higher in TW01 cells than those in DOK cells. (B) Bar graphs indicated a higher UBE2B/actin ratio in TW01 cells compared with that in DOK cells. (C) Expression levels of UBE2B in NPC cells treated with control or UBE2B-targeting siRNA were determined using western blot analysis. (D) By using methylene blue staining, the numbers of colonies formed, which consist of at least 50 tumor cells, were manually recorded and compared. (E) Bar graphs indicated decreased numbers of formed colonies in UBE2B-deficient NPC cells as compared with UBE2B-proficient cells. At least three independent experiments were performed and values were expressed as the mean ± standard deviation. ****P<0.01 for NPC cells vs. DOK cells and siUBE2B vs. control. Fold changes in protein levels listed under each blot were normalized to the levels of the control counterparts and analyzed by using ImageJ densitometry analysis. NPC, nasopharyngeal carcinoma; UBE2B, ubiquitin-conjugating enzyme E2 B; siRNA, small interfering RNA.

Article Snippet: The slides were then washed with tris-buffered saline and incubated with a primary rabbit polyclonal antibody against UBE2B (1:100 dilution; GeneTex, Inc.) for 1 h. A ChemMate EnVision kit (DAKO; Agilent Technologies, Inc.) was applied to detect the primary antibody.

Techniques: Western Blot, Expressing, Control, Staining, Standard Deviation, Ubiquitin Proteomics, Small Interfering RNA

Journal: Oncology Letters

Article Title: Role of high ubiquitin-conjugating enzyme E2 expression as a prognostic factor in nasopharyngeal carcinoma

doi: 10.3892/ol.2022.13314

Figure Lengend Snippet: UBE2B expression levels are a prognostic marker for patients with NPC receiving cisplatin-based chemoradiotherapy. Immunohistochemical analysis of UBE2B indicated nuclear and cytoplasmic staining in representative NPC cases with (A) low (H-score=125) and (B) high (H-score=375) expression (scale bars, 100 µm). (C) Survival analysis revealed that high expression of UBE2B was a prognostic marker for poor disease-specific survival, distal metastasis-free survival and local recurrence-free survival. NPC, nasopharyngeal carcinoma; UBE2B, ubiquitin-conjugating enzyme E2 B; Cum, cumulative.

Article Snippet: The slides were then washed with tris-buffered saline and incubated with a primary rabbit polyclonal antibody against UBE2B (1:100 dilution; GeneTex, Inc.) for 1 h. A ChemMate EnVision kit (DAKO; Agilent Technologies, Inc.) was applied to detect the primary antibody.

Techniques: Expressing, Marker, Immunohistochemical staining, Staining, Ubiquitin Proteomics

Journal: Oncology Letters

Article Title: Role of high ubiquitin-conjugating enzyme E2 expression as a prognostic factor in nasopharyngeal carcinoma

doi: 10.3892/ol.2022.13314

Figure Lengend Snippet: Associations between UBE2B expression levels and important clinicopathological variables.

Article Snippet: The slides were then washed with tris-buffered saline and incubated with a primary rabbit polyclonal antibody against UBE2B (1:100 dilution; GeneTex, Inc.) for 1 h. A ChemMate EnVision kit (DAKO; Agilent Technologies, Inc.) was applied to detect the primary antibody.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Role of high ubiquitin-conjugating enzyme E2 expression as a prognostic factor in nasopharyngeal carcinoma

doi: 10.3892/ol.2022.13314

Figure Lengend Snippet: Univariate log-rank analyses.

Article Snippet: The slides were then washed with tris-buffered saline and incubated with a primary rabbit polyclonal antibody against UBE2B (1:100 dilution; GeneTex, Inc.) for 1 h. A ChemMate EnVision kit (DAKO; Agilent Technologies, Inc.) was applied to detect the primary antibody.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Role of high ubiquitin-conjugating enzyme E2 expression as a prognostic factor in nasopharyngeal carcinoma

doi: 10.3892/ol.2022.13314

Figure Lengend Snippet: Multivariate survival analyses.

Article Snippet: The slides were then washed with tris-buffered saline and incubated with a primary rabbit polyclonal antibody against UBE2B (1:100 dilution; GeneTex, Inc.) for 1 h. A ChemMate EnVision kit (DAKO; Agilent Technologies, Inc.) was applied to detect the primary antibody.

Techniques: Expressing

Journal: Oncology Letters

Article Title: Role of high ubiquitin-conjugating enzyme E2 expression as a prognostic factor in nasopharyngeal carcinoma

doi: 10.3892/ol.2022.13314

Figure Lengend Snippet: UBE2B modulates cisplatin cytotoxicity in nasopharyngeal carcinoma cells by targeting MGMT expression. (A) Western blot analysis demonstrated UBE2B and MGMT expression in TW01 cells with distinctive siRNA or plasmid transfection. (B) Cell viability assays were performed with TW01 cells to analyze the role of UBE2B and MGMT in cisplatin-induced cell death by using methylene blue staining. At least three independent experiments were performed. Cell survival results were presented as the mean ± standard deviation and compared using analysis of variance with Tukey's post-hoc test. *P<0.01, ***P<0.0001, 2BKD group vs. control group; # P<0.01, ### P<0.0001, 2BKD + MGMT group vs. 2BKD group. Ctrl, cells transfected with scrambled siRNA; 2BKD, cells transfected with siUBE2B; 2BKD + MGMT, cells transfected with siUBE2B plus pEGFPc1-MGMT. UBE2B, ubiquitin-conjugating enzyme E2 B; siRNA, small interfering RNA; MGMT, O6-methylguanine-DNA methyltransferase; pEGFP, plasmid expressing enhanced green fluorescence protein.

Article Snippet: The slides were then washed with tris-buffered saline and incubated with a primary rabbit polyclonal antibody against UBE2B (1:100 dilution; GeneTex, Inc.) for 1 h. A ChemMate EnVision kit (DAKO; Agilent Technologies, Inc.) was applied to detect the primary antibody.

Techniques: Expressing, Western Blot, Plasmid Preparation, Transfection, Staining, Standard Deviation, Control, Ubiquitin Proteomics, Small Interfering RNA, Fluorescence