Journal: Frontiers in Immunology

Article Title: The role of APOBEC3C in modulating the tumor microenvironment and stemness properties of glioma: evidence from pancancer analysis

doi: 10.3389/fimmu.2023.1242972

Figure Lengend Snippet: Relationship between A3s expression and OS. (A) Expression of A3s identified as risk or protective factors in the TCGA and GTEx datasets. (B) Forest plot relating A3C expression to OS across 9 tumors.

Article Snippet: After blocking, the sections were incubated with an anti-A3C antibody (1:100; 10591-1-AP, Proteintech, USA) at 37°C for 1 h. The sections were washed 5 times with PBS and were then incubated with a secondary antibody (polyclonal rabbit IgG, Abcam).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: The role of APOBEC3C in modulating the tumor microenvironment and stemness properties of glioma: evidence from pancancer analysis

doi: 10.3389/fimmu.2023.1242972

Figure Lengend Snippet: Upregulation of A3C expression is associated with the prognosis and clinicopathological characteristics of glioma. (A) The gene–gene interaction network for the APOBEC3 family was constructed using the GeneMANIA database (B) The top 10 genes were identified by sorting the 18 genes encoding these proteins with supporting data from hub gene analysis. (C) Kaplan−Meier survival curves of patients in the TCGA-GBMLGG cohort based on A3C expression. The clinical features related to A3C expression included (D) tumor grade and (E) histology, based on analysis of the TCGA-GBMLGG cohort. (F) Kaplan–Meier survival curves of patients in the CGGA cohort based on A3C expression. The clinical features related to A3C expression included (G) tumor grade and (H) histology, based on analysis of the CGGA cohort. *P< 0.05, **P< 0.01, ***P< 0.001, ns, no significance.

Article Snippet: After blocking, the sections were incubated with an anti-A3C antibody (1:100; 10591-1-AP, Proteintech, USA) at 37°C for 1 h. The sections were washed 5 times with PBS and were then incubated with a secondary antibody (polyclonal rabbit IgG, Abcam).

Techniques: Expressing, Construct

Journal: Frontiers in Immunology

Article Title: The role of APOBEC3C in modulating the tumor microenvironment and stemness properties of glioma: evidence from pancancer analysis

doi: 10.3389/fimmu.2023.1242972

Figure Lengend Snippet: Relationships between A3C expression and glioma cell subpopulations. (A–C) A3C was expressed mainly in astrocytes, CD8+ T cells, and microglia in glioblastoma. (D) Associations between A3C expression and immune infiltration in GBM and LGG. (E) Associations between A3C expression and immune checkpoint expression in GBM and LGG. (F) Prognostic associations between A3C expression and immune infiltration in GBM and LGG. *P<0.05, **P< 0.01.

Article Snippet: After blocking, the sections were incubated with an anti-A3C antibody (1:100; 10591-1-AP, Proteintech, USA) at 37°C for 1 h. The sections were washed 5 times with PBS and were then incubated with a secondary antibody (polyclonal rabbit IgG, Abcam).

Techniques: Expressing

Journal: Frontiers in Immunology

Article Title: The role of APOBEC3C in modulating the tumor microenvironment and stemness properties of glioma: evidence from pancancer analysis

doi: 10.3389/fimmu.2023.1242972

Figure Lengend Snippet: A3C is significantly upregulated in glioma and is involved in maintenance of self-renewal in glial stem cells. (A) Boxplot showing A3C expression in different glioma tissues in the GEO database. (B) Expression level of A3C in different glioma cell lines (data from CCLE). (C) RT–qPCR results showing increased serum levels of A3C in GBM patients. (D) Evaluation of A3C expression by immunohistochemical staining in glioma patients. (E, F) GEPIA was used to show the correlations between A3 expression and stemness gene expression in GBM and LGG patients. (G, H) Representative images of neurosphere-cultured glial stem cells and two established glial stem cell lines. (I, J) Sphere formation of glioma cells was evaluated by the sphere formation assay. Knockdown of A3C inhibited sphere formation in glioma stem cells. T. Bar: 100 μm. (K,L) Knockdown of A3C inhibited stem cell marker expression by RT-qPCR. *P< 0.05, **P< 0.01, ***P< 0.001.

Article Snippet: After blocking, the sections were incubated with an anti-A3C antibody (1:100; 10591-1-AP, Proteintech, USA) at 37°C for 1 h. The sections were washed 5 times with PBS and were then incubated with a secondary antibody (polyclonal rabbit IgG, Abcam).

Techniques: Expressing, Quantitative RT-PCR, Immunohistochemical staining, Staining, Gene Expression, Cell Culture, Tube Formation Assay, Knockdown, Marker

Journal: Frontiers in Immunology

Article Title: The role of APOBEC3C in modulating the tumor microenvironment and stemness properties of glioma: evidence from pancancer analysis

doi: 10.3389/fimmu.2023.1242972

Figure Lengend Snippet: A3C knockdown impairs glioma cell proliferation, migration, and invasion. (A, B) Cell viability was tested by CCK8 assay. (C, D) Cell invasion was assessed by transwell migration invasion assays. (E, F) Cell migration was evaluated by wound healing assays. *P< 0.05, **P< 0.01, ***P< 0.001.

Article Snippet: After blocking, the sections were incubated with an anti-A3C antibody (1:100; 10591-1-AP, Proteintech, USA) at 37°C for 1 h. The sections were washed 5 times with PBS and were then incubated with a secondary antibody (polyclonal rabbit IgG, Abcam).

Techniques: Knockdown, Migration, CCK-8 Assay