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aoi wt cas9 (Addgene inc)

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Addgene inc aoi wt cas9
Aoi Wt Cas9, supplied by Addgene inc, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/aoi+wt+cas9/AOI-WT-Cas9-sg-mouse+Ezh2-E10-GFP+(Plasmid+%2391879)/pmc11121204-107-11-13
Average 92 stars, based on 1 article reviews

aoi wt cas9 - by Bioz Stars, 2026-09

92/100 stars

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Article Title: Tumor-Intrinsic Enhancer of Zeste Homolog 2 Controls Immune Cell Infiltration, Tumor Growth, and Lung Metastasis in a Triple-Negative Breast Cancer Model
Article Snippet: .. For KO plasmids, the murine EZH2 sequences from AOI-WT-Cas9-sq-mouse Ezh2-E18-GFP and AOI-WT-Cas9-sq-mouse Ezh2-E10-GFP (Addgene #91880 and Addgene #91879; gift from Martine Roussel [ ]) were cloned into pSPCas9(BB)-2A-Puro (Addgene #62988; gift from Feng Zhang [ ]). .. Plasmids were transfected into 4T1 cells using lipofectamine 3000 (Thermo Fisher Scientific) following the manufacturer’s protocol and selected using puromycin at a concentration of 5 μg/mL.

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EZH2 knockout (KO) and overexpression (OE) cell clones were generated from the parent wild-type (WT) 4T1 TNBC line. ( A ) Western blots (L indicates protein ladder) and ( B ) densitometric analysis comparing 11 clones from each EZH2 KO (upper panel) and EZH2 OE (lower panel) lines to the 4T1 WT line (2 sets of blots for each), with alpha-tubulin as the normalization control. ( C ) Comparison of EZH2 protein expression between KO (blue squares) and OE (red triangles) groups by Wilcoxon rank-sum test, **** p < 0.0001.

Journal: International Journal of Molecular Sciences

Article Title: Tumor-Intrinsic Enhancer of Zeste Homolog 2 Controls Immune Cell Infiltration, Tumor Growth, and Lung Metastasis in a Triple-Negative Breast Cancer Model

doi: 10.3390/ijms25105392

Figure Lengend Snippet: EZH2 knockout (KO) and overexpression (OE) cell clones were generated from the parent wild-type (WT) 4T1 TNBC line. ( A ) Western blots (L indicates protein ladder) and ( B ) densitometric analysis comparing 11 clones from each EZH2 KO (upper panel) and EZH2 OE (lower panel) lines to the 4T1 WT line (2 sets of blots for each), with alpha-tubulin as the normalization control. ( C ) Comparison of EZH2 protein expression between KO (blue squares) and OE (red triangles) groups by Wilcoxon rank-sum test, **** p < 0.0001.

Article Snippet: For KO plasmids, the murine EZH2 sequences from AOI-WT-Cas9-sq-mouse Ezh2-E18-GFP and AOI-WT-Cas9-sq-mouse Ezh2-E10-GFP (Addgene #91880 and Addgene #91879; gift from Martine Roussel [ ]) were cloned into pSPCas9(BB)-2A-Puro (Addgene #62988; gift from Feng Zhang [ ]).

Techniques: Knock-Out, Over Expression, Clone Assay, Generated, Western Blot, Control, Comparison, Expressing

In vitro replicative and invasive behaviors of EZH2 KO and EZH2 OE compared to parent WT 4T1 cells. ( A ) Counts of 4T1 WT, EZH2 KO, and EZH2 OE lines over 72 h of growth in 2D plates. ( B ) Representative images of spheroid for 4T1 WT, EZH2 KO, and EZH2 OE lines, and quantification of ( C ) circularity and ( D ) invasive area over 72 h of growth in a 3D invasion assay (n = 8 spheroids per group). Comparisons across groups and timepoints are by two-way ANOVA with Tukey’s post-hoc test and shown as * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Journal: International Journal of Molecular Sciences

Article Title: Tumor-Intrinsic Enhancer of Zeste Homolog 2 Controls Immune Cell Infiltration, Tumor Growth, and Lung Metastasis in a Triple-Negative Breast Cancer Model

doi: 10.3390/ijms25105392

Figure Lengend Snippet: In vitro replicative and invasive behaviors of EZH2 KO and EZH2 OE compared to parent WT 4T1 cells. ( A ) Counts of 4T1 WT, EZH2 KO, and EZH2 OE lines over 72 h of growth in 2D plates. ( B ) Representative images of spheroid for 4T1 WT, EZH2 KO, and EZH2 OE lines, and quantification of ( C ) circularity and ( D ) invasive area over 72 h of growth in a 3D invasion assay (n = 8 spheroids per group). Comparisons across groups and timepoints are by two-way ANOVA with Tukey’s post-hoc test and shown as * p < 0.05, ** p < 0.01, *** p < 0.001, and **** p < 0.0001.

Techniques: In Vitro, Invasion Assay

In vitro surface phenotype and secreted factors by EZH2 KO and EZH2 OE compared to parent WT 4T1 cells. ( A ) 4T1 WT (black circles), EZH2 KO (blue squares), and EZH2 OE (red triangles) lines were cultured in DMEM for 24 h and analyzed for the surface expression of relevant surface markers by flow cytometry (six repeats, see Methods and for details). ( B ) Culture supernatants were screened for relevant extracellular mediators via mesoscale assay (four repeats). Comparison between groups is by one-way ANOVA with Tukey’s post-hoc test and shown as * p < 0.05, ** p < 0.01.

Journal: International Journal of Molecular Sciences

Article Title: Tumor-Intrinsic Enhancer of Zeste Homolog 2 Controls Immune Cell Infiltration, Tumor Growth, and Lung Metastasis in a Triple-Negative Breast Cancer Model

doi: 10.3390/ijms25105392

Figure Lengend Snippet: In vitro surface phenotype and secreted factors by EZH2 KO and EZH2 OE compared to parent WT 4T1 cells. ( A ) 4T1 WT (black circles), EZH2 KO (blue squares), and EZH2 OE (red triangles) lines were cultured in DMEM for 24 h and analyzed for the surface expression of relevant surface markers by flow cytometry (six repeats, see Methods and for details). ( B ) Culture supernatants were screened for relevant extracellular mediators via mesoscale assay (four repeats). Comparison between groups is by one-way ANOVA with Tukey’s post-hoc test and shown as * p < 0.05, ** p < 0.01.

Techniques: In Vitro, Cell Culture, Expressing, Flow Cytometry, Comparison

In vivo primary tumor growth and lung metastasis by EZH2 KO and EZH2 OE compared to parent WT 4T1 cells. ( A ) Experimental timeline of 4T1 WT, EZH2 KO, and EZH2 OE injections in mice. ( B ) Growth of 4T1 WT, EZH2 KO, and EZH2 OE primary tumors over 21 days post-injection (n = 6–7 mice per group). Comparisons across groups and timepoints are by two-way ANOVA with Tukey’s post-hoc test and shown as * p < 0.05 and ** p < 0.01 (brackets). Comparison between groups at each timepoint is by one-way ANOVA and shown as * p < 0.05 (as indicated for WT vs. EZH2 KO and EZH2 KO vs. EZH2 OE, above each timepoint). ( C ) Lung metastasis assays for 4T1 WT, EZH2 KO, and EZH2 OE lines. Comparisons between groups are by one-way ANOVA with Tukey’s post-hoc test and shown as ** p < 0.01 and *** p < 0.001.

Journal: International Journal of Molecular Sciences

Article Title: Tumor-Intrinsic Enhancer of Zeste Homolog 2 Controls Immune Cell Infiltration, Tumor Growth, and Lung Metastasis in a Triple-Negative Breast Cancer Model

doi: 10.3390/ijms25105392

Figure Lengend Snippet: In vivo primary tumor growth and lung metastasis by EZH2 KO and EZH2 OE compared to parent WT 4T1 cells. ( A ) Experimental timeline of 4T1 WT, EZH2 KO, and EZH2 OE injections in mice. ( B ) Growth of 4T1 WT, EZH2 KO, and EZH2 OE primary tumors over 21 days post-injection (n = 6–7 mice per group). Comparisons across groups and timepoints are by two-way ANOVA with Tukey’s post-hoc test and shown as * p < 0.05 and ** p < 0.01 (brackets). Comparison between groups at each timepoint is by one-way ANOVA and shown as * p < 0.05 (as indicated for WT vs. EZH2 KO and EZH2 KO vs. EZH2 OE, above each timepoint). ( C ) Lung metastasis assays for 4T1 WT, EZH2 KO, and EZH2 OE lines. Comparisons between groups are by one-way ANOVA with Tukey’s post-hoc test and shown as ** p < 0.01 and *** p < 0.001.

Techniques: In Vivo, Injection, Comparison

In vivo primary tumor infiltration by neutrophils and CD4+ and CD8+ T cells for EZH2 KO and EZH2 OE compared to parent WT 4T1 cells. ( A ) Flow cytometry strategy for gating of infiltrating leukocyte subsets in 4T1 WT, EZH2 KO, and EZH2 OE primary tumors, with sequential steps 1 (leukocytes), 2 (live cells), 3 (singlets), 4 (granulocytes, including mature neutrophils, immature neutrophils, and eosinophils), 5 (non-granulocytes), 6 (T cells), and 7 (CD4+ and CD8+). ( B ) Relative percentages of infiltrating neutrophils and CD4+ and CD8+ cells among live leukocytes (top), and ratios between these subsets (bottom) in 4T1 WT (black circles), EZH2 KO (blue squares), and EZH2 OE (red triangles) primary tumors. Comparisons between groups are by one-way ANOVA with Tukey’s post-hoc test and shown as * p < 0.05 and ** p < 0.01.

Journal: International Journal of Molecular Sciences

Article Title: Tumor-Intrinsic Enhancer of Zeste Homolog 2 Controls Immune Cell Infiltration, Tumor Growth, and Lung Metastasis in a Triple-Negative Breast Cancer Model

doi: 10.3390/ijms25105392

Figure Lengend Snippet: In vivo primary tumor infiltration by neutrophils and CD4+ and CD8+ T cells for EZH2 KO and EZH2 OE compared to parent WT 4T1 cells. ( A ) Flow cytometry strategy for gating of infiltrating leukocyte subsets in 4T1 WT, EZH2 KO, and EZH2 OE primary tumors, with sequential steps 1 (leukocytes), 2 (live cells), 3 (singlets), 4 (granulocytes, including mature neutrophils, immature neutrophils, and eosinophils), 5 (non-granulocytes), 6 (T cells), and 7 (CD4+ and CD8+). ( B ) Relative percentages of infiltrating neutrophils and CD4+ and CD8+ cells among live leukocytes (top), and ratios between these subsets (bottom) in 4T1 WT (black circles), EZH2 KO (blue squares), and EZH2 OE (red triangles) primary tumors. Comparisons between groups are by one-way ANOVA with Tukey’s post-hoc test and shown as * p < 0.05 and ** p < 0.01.

Techniques: In Vivo, Flow Cytometry

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