Journal: Frontiers in Oncology

Article Title: WDR5 is a prognostic biomarker of brain metastasis from non-small cell lung cancer

doi: 10.3389/fonc.2022.1023776

Figure Lengend Snippet: Basic information of the inception and validation cohort.

Article Snippet: Primary antibody of WDR5 (R&D Systems, AF5810) was used to incubate the tissues at dilution of 1:50 overnight at 4°C, and the corresponding secondary antibody labelled with streptavidin-biotin-peroxidase reagent (Beyotime, Beijing, China) was applied to incubate the specimens for 30 minutes at room temperature.

Techniques: Biomarker Discovery

Journal: Frontiers in Oncology

Article Title: WDR5 is a prognostic biomarker of brain metastasis from non-small cell lung cancer

doi: 10.3389/fonc.2022.1023776

Figure Lengend Snippet: The expression of WDR5 in BM from NSCLC. The expression of WDR5 in BM from NSCLC was detected with IHC, dividing the cohort into high and low WDR5 expression. Black arrow indicates BM, and white arrow indicates brain tissue.

Article Snippet: Primary antibody of WDR5 (R&D Systems, AF5810) was used to incubate the tissues at dilution of 1:50 overnight at 4°C, and the corresponding secondary antibody labelled with streptavidin-biotin-peroxidase reagent (Beyotime, Beijing, China) was applied to incubate the specimens for 30 minutes at room temperature.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: WDR5 is a prognostic biomarker of brain metastasis from non-small cell lung cancer

doi: 10.3389/fonc.2022.1023776

Figure Lengend Snippet: The correlation between WDR5 and the clinicopathological factors.

Article Snippet: Primary antibody of WDR5 (R&D Systems, AF5810) was used to incubate the tissues at dilution of 1:50 overnight at 4°C, and the corresponding secondary antibody labelled with streptavidin-biotin-peroxidase reagent (Beyotime, Beijing, China) was applied to incubate the specimens for 30 minutes at room temperature.

Techniques:

Journal: Frontiers in Oncology

Article Title: WDR5 is a prognostic biomarker of brain metastasis from non-small cell lung cancer

doi: 10.3389/fonc.2022.1023776

Figure Lengend Snippet: Prognostic significance in univariate and multivariate analyses.

Article Snippet: Primary antibody of WDR5 (R&D Systems, AF5810) was used to incubate the tissues at dilution of 1:50 overnight at 4°C, and the corresponding secondary antibody labelled with streptavidin-biotin-peroxidase reagent (Beyotime, Beijing, China) was applied to incubate the specimens for 30 minutes at room temperature.

Techniques:

Journal: Frontiers in Oncology

Article Title: WDR5 is a prognostic biomarker of brain metastasis from non-small cell lung cancer

doi: 10.3389/fonc.2022.1023776

Figure Lengend Snippet: Prognostic significance of WDR5 and clinicopathological factors. The cohort was divided into subsets according to WDR5 expression and clinicopathological factors. High WDR5 (A) and low KPS (B) were significantly associated with low OS rate. Patients’ age (C) , gender (D) , tumor size (E) and histological type (F) had no statistically significant correlation with OS of BM.

Article Snippet: Primary antibody of WDR5 (R&D Systems, AF5810) was used to incubate the tissues at dilution of 1:50 overnight at 4°C, and the corresponding secondary antibody labelled with streptavidin-biotin-peroxidase reagent (Beyotime, Beijing, China) was applied to incubate the specimens for 30 minutes at room temperature.

Techniques: Expressing

Journal: Oral surgery, oral medicine, oral pathology and oral radiology

Article Title: Osteogenic genes related to the canonic WNT pathway are down-regulated in ameloblastoma.

doi: 10.1016/j.oooo.2012.08.453

Figure Lengend Snippet: Fig. 1. Representative cases of immunohistochemistry of ameloblastoma showing immunostaining for (A, B) WDR5, (C, D) sFRP-2, and (E, F) RUNX2. (G, H) Representative cases showing nuclear -catenin–positive cells (arrows).

Article Snippet: Immunohistochemical staining was done as previously described.19 The primary antibodies used were mouse monoclonal WDR5 (Santa Cruz Bio- tech, Santa Cruz, CA; diluted 1:100) and RUNX2 (Abcam, Cambridge, MA; diluted 1:150), rabbit polyclonal sFRP-2 (Santa Cruz Biotech, Santa Cruz, CA; diluted 1:300), and -catenin (Dako, Copenhagen, Denmark; diluted 1:200).

Techniques: Immunohistochemistry, Immunostaining

Journal: Oral surgery, oral medicine, oral pathology and oral radiology

Article Title: Osteogenic genes related to the canonic WNT pathway are down-regulated in ameloblastoma.

doi: 10.1016/j.oooo.2012.08.453

Figure Lengend Snippet: Fig. 2. A, WDR5 and sFRP-2 proteins were detected in total cell lysates extracted from AM-1 cells by Western blot analysis. Tubulin was shown as a protein loading control of total cell lysates. B, mRNA expression of RUNX2, C-MYC, and WDR5 was detected in AM-1 cells; C, mRNA expression of RUNX2 and C-MYC was detected in KUSA/A1 cells. In both type of cells, GAPDH was shown as a control gene. D, The relative expression levels in KUSA/A1 cells were calculated and data were normalized to the GAPDH relative to experimental gene expression, which represents fold change. Asterisks indicate P .05.

Article Snippet: Immunohistochemical staining was done as previously described.19 The primary antibodies used were mouse monoclonal WDR5 (Santa Cruz Bio- tech, Santa Cruz, CA; diluted 1:100) and RUNX2 (Abcam, Cambridge, MA; diluted 1:150), rabbit polyclonal sFRP-2 (Santa Cruz Biotech, Santa Cruz, CA; diluted 1:300), and -catenin (Dako, Copenhagen, Denmark; diluted 1:200).

Techniques: Western Blot, Control, Expressing, Gene Expression

Journal: Oral surgery, oral medicine, oral pathology and oral radiology

Article Title: Osteogenic genes related to the canonic WNT pathway are down-regulated in ameloblastoma.

doi: 10.1016/j.oooo.2012.08.453

Figure Lengend Snippet: Fig. 3. Western blot analysis of KUSA/A1 cells cultured in 10-cm culture plates in OSM, AM-1CM, or 1ksfm:2osm. A, After confluent Western blot was performed. WDR5 protein was detected in total cell lysates extracted from KUSA/A1 cells by Western blot analysis. Tubulin was shown as a protein loading control of total cell lysates. Dose-dependent induction of alkaline phosphatase activity was conducted by recombinant WDR5 protein and compared with the cells cultured with AM-1CM or OSM. B, On the next day after seeding, cells were treated with the protein for 48 hours, followed by assessment of alkaline phosphatase activity and data from absorbance was subjected to statistical analysis. Asterisks indicate P .05.

Article Snippet: Immunohistochemical staining was done as previously described.19 The primary antibodies used were mouse monoclonal WDR5 (Santa Cruz Bio- tech, Santa Cruz, CA; diluted 1:100) and RUNX2 (Abcam, Cambridge, MA; diluted 1:150), rabbit polyclonal sFRP-2 (Santa Cruz Biotech, Santa Cruz, CA; diluted 1:300), and -catenin (Dako, Copenhagen, Denmark; diluted 1:200).

Techniques: Western Blot, Cell Culture, Control, Activity Assay, Recombinant

Journal: Epigenetics & chromatin

Article Title: Roles for common MLL/COMPASS subunits and the 19S proteasome in regulating CIITA pIV and MHC class II gene expression and promoter methylation.

doi: 10.1186/1756-8935-3-5

Figure Lengend Snippet: Figure 2 Overexpressing a single common subunit of MLL/COMPASS complexes enhances CIITA transactivity. (A-C) Overexpression of common MLL/COMPASS subunits increases CIITA transactivity at the MHC-II promoter. HeLa cells were transfected with HLA-DRA-Luc, Renilla, pcDNA3, CIITA and (a) WDR5, (b) Ash2L or (c) RbBP5. Luciferase activity is reported as fold activation relative to that of the reporter alone. Luciferase readings were normalized to Renilla activity. Assays were performed in triplicate and values represent mean ± SEM of (n = 3) independent experiments. (d-e) Overexpression of common MLL/COMPASS subunits increases MHC-II transcript levels. HeLa cells were either left untransfected or were transfected with CIITA alone or in combination with WDR5, Ash2L or RbBP5. RNA was prepared using TRIzol reagent and cDNA was generated using gene specific antisense primers for (d) MHC-II HLA-DR and (e) GAPDH. cDNA was quantified via real-time PCR, and data are graphed as fold changes over non-transfected samples. Values represent mean ± SEM of (n = 2) independent experiments. *P < 0.05 vs control siRNA.

Article Snippet: Pooled siRNA duplexes for WDR5 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Over Expression, Transfection, Luciferase, Activity Assay, Activation Assay, Generated, Real-time Polymerase Chain Reaction, Control

Journal: Epigenetics & chromatin

Article Title: Roles for common MLL/COMPASS subunits and the 19S proteasome in regulating CIITA pIV and MHC class II gene expression and promoter methylation.

doi: 10.1186/1756-8935-3-5

Figure Lengend Snippet: Figure 3 Knockdown of a common MLL/COMPASS subunit decreases H3K4me3. (a) WDR5 knockdown efficiently reduces WDR5 expression. HeLa cells were transfected with control or WDR5-specific siRNA, harvested and subjected to western blot analysis for (top) endogenous WDR5 and (bottom) endogenous tubulin. (b-c) WDR5 knockdown decreases H3K4me3. HeLa cells transfected with scrambled control or WDR5-specific siRNA were stimulated with IFN-g and subjected to ChIP assay. Lysates were immunoprecipitated with control or endogenous H3K4me3 antibody. Associated DNA was isolated and analyzed via real-time PCR as described in Figure 1, using primers and probes specific for (b) the MHC-II promoter or (c) CIITA pIV. Data are presented as percentage input. IgG Isotype control values were 0.06 ± 0.03 (MHC-II promoter) and 0.003 ± 0.002 (CIITA pIV). Values represent mean ± SEM of (n = 3) independent experiments. *P < 0.05 vs control siRNA.

Article Snippet: Pooled siRNA duplexes for WDR5 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Knockdown, Expressing, Transfection, Control, Western Blot, Immunoprecipitation, Isolation, Real-time Polymerase Chain Reaction

Journal: Epigenetics & chromatin

Article Title: Roles for common MLL/COMPASS subunits and the 19S proteasome in regulating CIITA pIV and MHC class II gene expression and promoter methylation.

doi: 10.1186/1756-8935-3-5

Figure Lengend Snippet: Figure 4 WDR5 knockdown does not affect H3K18ac. (a, b) HeLa cells transfected with scrambled control or WDR5-specific siRNA were stimulated with IFN-g and subjected to ChIP assay. Lysates were immunoprecipitated with control or endogenous H3K18ac antibody. Associated DNA was isolated and analyzed via real-time PCR as described in Figure 2, using primers and probes specific for (a) MHC-II promoter or (b) CIITA pIV. Data are presented as percentage input. IgG Isotype control values were 0.06 ± 0.03 (MHC-II promoter) and 0.003 ± 0.002 (CIITA pIV). Values represent mean ± SEM of (n = 3) independent experiments.

Article Snippet: Pooled siRNA duplexes for WDR5 were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Knockdown, Transfection, Control, Immunoprecipitation, Isolation, Real-time Polymerase Chain Reaction