Journal: Scientific Reports

Article Title: Deciphering the prognostic significance of WDR77 in gliomas: a comprehensive analysis

doi: 10.1038/s41598-024-82867-w

Figure Lengend Snippet: Expression analysis of WDR77 in glioma datasets and tissue Samples. ( A ) The expression of WDR77 in different glioma datasets. ( B ) Expression detection of WDR77 in glioma samples. ( C ) Grayscale statistical results of WDR77 expression in different grades of glioma tissue relative to GAPDH. ( D ) WDR77 expression in GBM cell lines U251 by IHC. p < 0.01 is considered statistically significant.

Article Snippet: Furthermore, we queried the Human Protein Atlas (HPA) database ( https://www.proteinatlas.org/ ) for WDR77 expression in the GBM cell line U251.

Techniques: Expressing, Paraffin-embedded Immunohistochemistry

Journal: Scientific Reports

Article Title: Deciphering the prognostic significance of WDR77 in gliomas: a comprehensive analysis

doi: 10.1038/s41598-024-82867-w

Figure Lengend Snippet: Correlation between WDR77 expression and glioma molecular subtypes. ( A – D ) WDR77 was significantly enriched in the 1p19q non-codel subtype from the TCGA and CGGA cohorts. ( B – E ) WDR77 was significantly enriched in the IDH wild type subtype from the TCGA and CGGA cohorts . ( C – F ) The expression of WDR77 was not statistically different between male and female glioma patients. ( G ) Among the oligoderdroglioma, oligoastrocytoma and GBM subtypes, WDR77 is the most expressed in GBM. (H–I) ROC curve analysis showing the predictive value of WDR77 in the TCGA and CGGA cohorts. *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: Furthermore, we queried the Human Protein Atlas (HPA) database ( https://www.proteinatlas.org/ ) for WDR77 expression in the GBM cell line U251.

Techniques: Expressing

Journal: Scientific Reports

Article Title: Deciphering the prognostic significance of WDR77 in gliomas: a comprehensive analysis

doi: 10.1038/s41598-024-82867-w

Figure Lengend Snippet: Survival curve of WDR77 in different databases. ( A ) Survival curve of WDR77 in Rembrandt database. ( B ) Survival curve of WDR77 in Gravendeel databset. ( C ) Survival curve of WDR77 in the TCGA database. ( D ) Survival curve of WDR77 in CGGA database. ( E ) Univariate Cox analyses evaluating the independent prognostic value of WDR77 in glioma patients from the CGGA database. ( F ) Multivariate Cox analyses evaluating the independent prognostic value of WDR77 in glioma patients from the CGGA database.

Article Snippet: Furthermore, we queried the Human Protein Atlas (HPA) database ( https://www.proteinatlas.org/ ) for WDR77 expression in the GBM cell line U251.

Techniques:

Journal: Scientific Reports

Article Title: Deciphering the prognostic significance of WDR77 in gliomas: a comprehensive analysis

doi: 10.1038/s41598-024-82867-w

Figure Lengend Snippet: Effect of WDR77 knockdown in glioma cells. ( A ) The relationship between WDR77 and cell cycle function-related genes in TCGA glioma samples. ( B ) Flow cytometry analysis of cell cycle distribution in U87-MG and U251 glioma cells. Cells were treated with different concentrations (5 µmol, 10 µmol, 15 µmol) of siRNA targeting WDR77 . WT represents wild-type, untreated cells. The x-axis shows the DNA content, and the y-axis shows the cell count. The peaks correspond to the G1, S, and G2 phases of the cell cycle. ( C ) Bar graphs showing the percentage distribution of cells in the G1, S, and G2 phases for U87-MG and U251 cells treated with different concentrations of siRNA-WDR77 (5 µmol, 10 µmol, 20 µmol).

Article Snippet: Furthermore, we queried the Human Protein Atlas (HPA) database ( https://www.proteinatlas.org/ ) for WDR77 expression in the GBM cell line U251.

Techniques: Knockdown, Flow Cytometry, Cell Counting

Journal: Scientific Reports

Article Title: Deciphering the prognostic significance of WDR77 in gliomas: a comprehensive analysis

doi: 10.1038/s41598-024-82867-w

Figure Lengend Snippet: Correlation between WDR77 Levels and Immune Cell Infiltration in Glioma. ( A ) The correlation between WDR77 levels and immune cell-specific marker genes using the online analysis tool TIMER 2.0. ( B , C ) We evaluated the association between the expression of common immune infiltrating cell-specific markers (Treg cells, Macrophage cells, Dendritic cells, and B cells) and the WDR77 gene. Our analysis established that WDR77 levels were positively correlated with B cells (GBM: R = 0.409, p < 0.01; LGG: R = − 0.193, p < 0.01), Treg cells (GBM: R = 0.409, p < 0.01; LGG: R = − 0.193, p < 0.01), Macrophage cells (GBM: R = 0.409, p < 0.01; LGG: R = 0.409, p < 0.01), Dendritic cells (GBM: R = 0.409, p < 0.01; LGG: R = − 0.193, p < 0.01). These results suggest that glioma tissues with elevated WDR77 levels are highly infiltrated with invasive immune cells, especially those with immunosuppressive characteristics.

Article Snippet: Furthermore, we queried the Human Protein Atlas (HPA) database ( https://www.proteinatlas.org/ ) for WDR77 expression in the GBM cell line U251.

Techniques: Marker, Expressing

Journal: Scientific Reports

Article Title: Deciphering the prognostic significance of WDR77 in gliomas: a comprehensive analysis

doi: 10.1038/s41598-024-82867-w

Figure Lengend Snippet: ( A – V ) The WDR77 interaction network of co-expressed genes in different tumours. The coexpression network was drawn using R package, Only the top 20 genes with the highest correlations are shown. Red circle shows input gene, orange circle represents cell metabolism gene, and the sky circle represents other genes.

Article Snippet: Furthermore, we queried the Human Protein Atlas (HPA) database ( https://www.proteinatlas.org/ ) for WDR77 expression in the GBM cell line U251.

Techniques:

Journal: Cancers

Article Title: PRMT5/WDR77 Enhances the Proliferation of Squamous Cell Carcinoma via the ΔNp63α-p21 Axis.

doi: 10.3390/cancers16223789

Figure Lengend Snippet: Figure 1. PRMT5 is inversely correlated with SCC survival. (A) Statistical analysis of the TCGA- HNSC database comparing the expression of PRMT5 (left) and WDR77 (right) in cancer (red) vs. normal (blue) groups, utilizing Wilcoxon Rank-Sum Tests. Patient counts: HNSCC (513 tumors and 44 normal). Black dots represent median expression levels. (B) Kaplan–Meier survival curve for PRMT5 and WDR77 expression in the TCGA-HNSC database (n = 513). Patients were categorized by median PRMT5 (upper panel) and WDR77 (lower panel) expression levels. Statistical significance was determined using the Log-Rank Tests, with visualization provided by the ggsurvplot function from the survminer R package [29]. (C) Statistical analysis of DepMap data and display of Chronos Dependency Scores for six subtypes of SCC. Chronos Dependency Scores, derived from cell depletion assays, indicate gene essentiality, where lower (more negative) scores reflect higher gene essentiality. (D) Immunohistochemical staining for PRMT5 (left) and ∆Np63α (right) in HNSCC patient specimens. The top panels show low-magnification views. Scale bars: 400 µm (top) and 100 µm (bottom).

Article Snippet: The list of antibodies used in this study includes: PRMT5 (Cell Signaling, Danvers, MA, USA, Cat. No. 79998), WDR77 (Cell Signaling, Danvers, MA, USA, Cat. No. 2823), ∆Np63α (Cell Signaling, Danvers, MA, USA, Cat. No. 39692), p21 (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-71811), β-Actin (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-47778), and HSC70 (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-7298).

Techniques: Expressing, Derivative Assay, Immunohistochemical staining, Staining

Journal: Cancers

Article Title: PRMT5/WDR77 Enhances the Proliferation of Squamous Cell Carcinoma via the ΔNp63α-p21 Axis.

doi: 10.3390/cancers16223789

Figure Lengend Snippet: Figure 2. PRMT5 and WDR77 regulate the HNSCC-specific transcriptome. (A) GSEA plots identified genes affected in both PRMT5KO and WDR77KO, with notably enriched RICK- MAN_HEAD_AND_NECK_CANCER C and E gene sets. (B) Heatmaps of GSEA results for both PRMT5KO and WDR77KO, highlighting the top five positively and negatively enriched gene sets. (C) UMAP visualizations depict cell types for each cluster and expression patterns of PRMT5, WDR77, and TP63. (D) Comparison of PRMT5 (left) or WDR77 (right), along with TP63, CDKN1A, and MKI67 gene expression, in PRMT5High vs. PRMT5Low (left) and in WDR77High vs. WDR77Low (right) subgroups in single HNSCC cells. p-values were calculated using the Wilcoxon Rank-Sum Tests, and the median (50th percentile) for each dataset is denoted by a solid line.

Article Snippet: The list of antibodies used in this study includes: PRMT5 (Cell Signaling, Danvers, MA, USA, Cat. No. 79998), WDR77 (Cell Signaling, Danvers, MA, USA, Cat. No. 2823), ∆Np63α (Cell Signaling, Danvers, MA, USA, Cat. No. 39692), p21 (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-71811), β-Actin (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-47778), and HSC70 (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-7298).

Techniques: Expressing, Comparison, Gene Expression

Journal: Cancers

Article Title: PRMT5/WDR77 Enhances the Proliferation of Squamous Cell Carcinoma via the ΔNp63α-p21 Axis.

doi: 10.3390/cancers16223789

Figure Lengend Snippet: Figure 3. Depletion of PRMT5 or WDR77 inhibits cell proliferation. (A) CRISPR-mediated depletion of PRMT5 or WDR77 in SCC cells using four sgRNAs (two for each gene). (B) Cellular competition-based GFP dropout assays in HSC5, FaDu, and Cal33 cells treated with sgRosa26, sgCDK1, sgPRMT5-1, sgPRMT5-2, sgWDR77-1, and sgWDR77-2. Normalized to P0. (C) MTT-based proliferation assays in the indicated SCC cell lines. Cells were treated with DMSO or the PRMT5 inhibitor PF-06939999 (4 µmol/L) for 48 h. Data were normalized to DMSO controls (n = 3 biologically independent replicates). The p-values were calculated using two-tailed unpaired Student’s t-tests. (D) Flow cytometry of HSC5 cells treated with sgNeg, sgPRMT5-1, and sgWDR77-2 (n = 3 biologically independent samples). The p-values were calculated using Two-Way ANOVA.

Article Snippet: The list of antibodies used in this study includes: PRMT5 (Cell Signaling, Danvers, MA, USA, Cat. No. 79998), WDR77 (Cell Signaling, Danvers, MA, USA, Cat. No. 2823), ∆Np63α (Cell Signaling, Danvers, MA, USA, Cat. No. 39692), p21 (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-71811), β-Actin (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-47778), and HSC70 (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-7298).

Techniques: CRISPR, Two Tailed Test, Flow Cytometry

Journal: Cancers

Article Title: PRMT5/WDR77 Enhances the Proliferation of Squamous Cell Carcinoma via the ΔNp63α-p21 Axis.

doi: 10.3390/cancers16223789

Figure Lengend Snippet: Figure 4. PRMT5 and WDR77 modulate the ∆Np63α-p21 pathway. (A) qRT-PCR for PRMT5, WDR77 and TP63 transcript levels in FaDu cells transduced with sgPRMT5-1 and sgWDR77-2, normalized to GAPDH. The p-values were calculated using One-Way ANOVA. (B) Western blot of PRMT5, WDR77, ∆Np63α, and p21 expression in FaDu cells transduced with sgNeg (empty vector control), sgPRMT5-1, sgPRMT5-2, sgWDR77-1, and sgWDR77-2. (C) FaDu cells were infected with sgNeg, sgPRMT5-1, and sgWDR77-2, with or without additional treatment of 1 µmol/L lactacystin for 24 h, as described [36]. (D) MTT-based proliferation assays in FaDu scramble cells (control) and FaDu cells overexpressing CRISPR-resistant ∆Np63α cDNAs (∆Np63α CR). Cells were infected with sgNeg, sgPRMT5-1, and sgWDR77-2. Data are presented as means ± S.D., normalized to sgNeg (n = 3 biologically independent samples). The p-values were calculated using One-Way ANOVA. (E) Confirmation of PRMT5KO and WDR77KO via Western blot in FaDu scramble cells (control) and FaDu cells overexpressing ∆Np63α CR. Cells were treated with sgNeg, sgPRMT5-1, and sgWDR77-2. (F) Assessment of PRMT5, WDR77, ∆Np63α, and p21 expression via Western blot in FaDu scramble cells (control) and FaDu cells overexpressing ∆Np63α CR. Cells were treated with sgNeg (empty vector control), sgp63-1, and sgp63-2. The uncropped bolts were shown in Supplementary Figure S7.

Article Snippet: The list of antibodies used in this study includes: PRMT5 (Cell Signaling, Danvers, MA, USA, Cat. No. 79998), WDR77 (Cell Signaling, Danvers, MA, USA, Cat. No. 2823), ∆Np63α (Cell Signaling, Danvers, MA, USA, Cat. No. 39692), p21 (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-71811), β-Actin (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-47778), and HSC70 (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-7298).

Techniques: Quantitative RT-PCR, Transduction, Western Blot, Expressing, Plasmid Preparation, Control, Infection, CRISPR

Journal: Cancers

Article Title: PRMT5/WDR77 Enhances the Proliferation of Squamous Cell Carcinoma via the ΔNp63α-p21 Axis.

doi: 10.3390/cancers16223789

Figure Lengend Snippet: Figure 5. PRMT5 or WDR77 depletion inhibits tumor growth in vivo. (A,B) Tumor volume caliper measurements and tumor weight measurements are means ± S.D. (n = 8). FaDu cells transduced with sgNeg, sgPRMT5-1, and sgWDR77-2 were injected subcutaneously into both rear flanks of nude mice (n = 4), with 5 × 104 cells per injection. Tumors were measured, weighed, and collected after 22 days. Mice were initially labeled as groups 1, 2, and 3, with the treatment groups blinded to ensure unbiased measurements until all data collection was completed. The p-values were calculated using One-Way ANOVA. (C) Representative images of tumors from each group. (D) Western blot assessment of PRMT5, WDR77, ∆Np63α, and p21 expression in the tumor samples from mice. The uncropped bolts were shown in Supplementary Figure S7.

Article Snippet: The list of antibodies used in this study includes: PRMT5 (Cell Signaling, Danvers, MA, USA, Cat. No. 79998), WDR77 (Cell Signaling, Danvers, MA, USA, Cat. No. 2823), ∆Np63α (Cell Signaling, Danvers, MA, USA, Cat. No. 39692), p21 (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-71811), β-Actin (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-47778), and HSC70 (Santa Cruz Biotechnology, Dallas, TX, USA, Cat. No. sc-7298).

Techniques: In Vivo, Transduction, Injection, Labeling, Western Blot, Expressing