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Figure 6. Deletion of Pericyte-b3-Integrin Enhances Tumor Cell Growth via Pericyte-pAkt-p65-Regulated Cytokine Production (A) Western blot analysis of b3-null pericytes with CRISPR-Cas9 deletion of <t>Akt1</t> (b3-null;AktKO PC) or HGFR (b3-null;HGFRKO PC) compared with b3-null;Cas9 controls. Hsc70, loading control. Blots representative of 3 separate experiments. Bar charts, average densitometric analysis relative to loading controls (n = 3 experimental repeats). (B) B16F0 cell survival after exposure to conditioned media from b3-null;AktKO, b3-null;HGFRKO, or b3-null;Cas9 pericytes (n = 16 technical replicates for 2 separate experiments). (C) Tumor growth in WT mice injected subcutaneously with either B16F0 cells plus b3-null;Cas9 pericytes or B16F0 plus b3-null;AktKO pericytes at 1:8 ratio (n = 10 per group). (D) Western blot analysis using mock- and Ik-Ba-SR-transfected WT or b3-null pericyte lysates. HSC70, loading control. (E) Protein-profiler cytokine arrays. Quantitation of fold difference in cytokine expression between mock- and Ik-Ba-SR-transfected b3-null pericytes (n = 4 dots from 2 independent experiments). (F) Conditioned media from mock- and Ik-Ba-SR-transfected b3-null pericytes were applied to B16F0, B16F10, and LLC cells and cell survival measured by MTS assay. Bar charts, means ± SEM relative to tumor cells treated with conditioned media from mock-transfected b3-null pericytes (n = 3 experimental repeats).
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Figure 6. Deletion of Pericyte-b3-Integrin Enhances Tumor Cell Growth via Pericyte-pAkt-p65-Regulated Cytokine Production (A) Western blot analysis of b3-null pericytes with CRISPR-Cas9 deletion of <t>Akt1</t> (b3-null;AktKO PC) or HGFR (b3-null;HGFRKO PC) compared with b3-null;Cas9 controls. Hsc70, loading control. Blots representative of 3 separate experiments. Bar charts, average densitometric analysis relative to loading controls (n = 3 experimental repeats). (B) B16F0 cell survival after exposure to conditioned media from b3-null;AktKO, b3-null;HGFRKO, or b3-null;Cas9 pericytes (n = 16 technical replicates for 2 separate experiments). (C) Tumor growth in WT mice injected subcutaneously with either B16F0 cells plus b3-null;Cas9 pericytes or B16F0 plus b3-null;AktKO pericytes at 1:8 ratio (n = 10 per group). (D) Western blot analysis using mock- and Ik-Ba-SR-transfected WT or b3-null pericyte lysates. HSC70, loading control. (E) Protein-profiler cytokine arrays. Quantitation of fold difference in cytokine expression between mock- and Ik-Ba-SR-transfected b3-null pericytes (n = 4 dots from 2 independent experiments). (F) Conditioned media from mock- and Ik-Ba-SR-transfected b3-null pericytes were applied to B16F0, B16F10, and LLC cells and cell survival measured by MTS assay. Bar charts, means ± SEM relative to tumor cells treated with conditioned media from mock-transfected b3-null pericytes (n = 3 experimental repeats).
Paper N A Plvpt Ttr Krab Szulc, 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
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Addgene inc human ttr precursor cdna
Figure 6. Deletion of Pericyte-b3-Integrin Enhances Tumor Cell Growth via Pericyte-pAkt-p65-Regulated Cytokine Production (A) Western blot analysis of b3-null pericytes with CRISPR-Cas9 deletion of <t>Akt1</t> (b3-null;AktKO PC) or HGFR (b3-null;HGFRKO PC) compared with b3-null;Cas9 controls. Hsc70, loading control. Blots representative of 3 separate experiments. Bar charts, average densitometric analysis relative to loading controls (n = 3 experimental repeats). (B) B16F0 cell survival after exposure to conditioned media from b3-null;AktKO, b3-null;HGFRKO, or b3-null;Cas9 pericytes (n = 16 technical replicates for 2 separate experiments). (C) Tumor growth in WT mice injected subcutaneously with either B16F0 cells plus b3-null;Cas9 pericytes or B16F0 plus b3-null;AktKO pericytes at 1:8 ratio (n = 10 per group). (D) Western blot analysis using mock- and Ik-Ba-SR-transfected WT or b3-null pericyte lysates. HSC70, loading control. (E) Protein-profiler cytokine arrays. Quantitation of fold difference in cytokine expression between mock- and Ik-Ba-SR-transfected b3-null pericytes (n = 4 dots from 2 independent experiments). (F) Conditioned media from mock- and Ik-Ba-SR-transfected b3-null pericytes were applied to B16F0, B16F10, and LLC cells and cell survival measured by MTS assay. Bar charts, means ± SEM relative to tumor cells treated with conditioned media from mock-transfected b3-null pericytes (n = 3 experimental repeats).
Human Ttr Precursor Cdna, supplied by Addgene inc, used in various techniques. Bioz Stars score: 93/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Addgene inc inducible system vector plvet ttr krab
Figure 6. Deletion of Pericyte-b3-Integrin Enhances Tumor Cell Growth via Pericyte-pAkt-p65-Regulated Cytokine Production (A) Western blot analysis of b3-null pericytes with CRISPR-Cas9 deletion of <t>Akt1</t> (b3-null;AktKO PC) or HGFR (b3-null;HGFRKO PC) compared with b3-null;Cas9 controls. Hsc70, loading control. Blots representative of 3 separate experiments. Bar charts, average densitometric analysis relative to loading controls (n = 3 experimental repeats). (B) B16F0 cell survival after exposure to conditioned media from b3-null;AktKO, b3-null;HGFRKO, or b3-null;Cas9 pericytes (n = 16 technical replicates for 2 separate experiments). (C) Tumor growth in WT mice injected subcutaneously with either B16F0 cells plus b3-null;Cas9 pericytes or B16F0 plus b3-null;AktKO pericytes at 1:8 ratio (n = 10 per group). (D) Western blot analysis using mock- and Ik-Ba-SR-transfected WT or b3-null pericyte lysates. HSC70, loading control. (E) Protein-profiler cytokine arrays. Quantitation of fold difference in cytokine expression between mock- and Ik-Ba-SR-transfected b3-null pericytes (n = 4 dots from 2 independent experiments). (F) Conditioned media from mock- and Ik-Ba-SR-transfected b3-null pericytes were applied to B16F0, B16F10, and LLC cells and cell survival measured by MTS assay. Bar charts, means ± SEM relative to tumor cells treated with conditioned media from mock-transfected b3-null pericytes (n = 3 experimental repeats).
Inducible System Vector Plvet Ttr Krab, 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
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Boster Bio rabbit pab
Figure 6. Deletion of Pericyte-b3-Integrin Enhances Tumor Cell Growth via Pericyte-pAkt-p65-Regulated Cytokine Production (A) Western blot analysis of b3-null pericytes with CRISPR-Cas9 deletion of <t>Akt1</t> (b3-null;AktKO PC) or HGFR (b3-null;HGFRKO PC) compared with b3-null;Cas9 controls. Hsc70, loading control. Blots representative of 3 separate experiments. Bar charts, average densitometric analysis relative to loading controls (n = 3 experimental repeats). (B) B16F0 cell survival after exposure to conditioned media from b3-null;AktKO, b3-null;HGFRKO, or b3-null;Cas9 pericytes (n = 16 technical replicates for 2 separate experiments). (C) Tumor growth in WT mice injected subcutaneously with either B16F0 cells plus b3-null;Cas9 pericytes or B16F0 plus b3-null;AktKO pericytes at 1:8 ratio (n = 10 per group). (D) Western blot analysis using mock- and Ik-Ba-SR-transfected WT or b3-null pericyte lysates. HSC70, loading control. (E) Protein-profiler cytokine arrays. Quantitation of fold difference in cytokine expression between mock- and Ik-Ba-SR-transfected b3-null pericytes (n = 4 dots from 2 independent experiments). (F) Conditioned media from mock- and Ik-Ba-SR-transfected b3-null pericytes were applied to B16F0, B16F10, and LLC cells and cell survival measured by MTS assay. Bar charts, means ± SEM relative to tumor cells treated with conditioned media from mock-transfected b3-null pericytes (n = 3 experimental repeats).
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Addgene inc lentiviral vector 32
Figure 6. Deletion of Pericyte-b3-Integrin Enhances Tumor Cell Growth via Pericyte-pAkt-p65-Regulated Cytokine Production (A) Western blot analysis of b3-null pericytes with CRISPR-Cas9 deletion of <t>Akt1</t> (b3-null;AktKO PC) or HGFR (b3-null;HGFRKO PC) compared with b3-null;Cas9 controls. Hsc70, loading control. Blots representative of 3 separate experiments. Bar charts, average densitometric analysis relative to loading controls (n = 3 experimental repeats). (B) B16F0 cell survival after exposure to conditioned media from b3-null;AktKO, b3-null;HGFRKO, or b3-null;Cas9 pericytes (n = 16 technical replicates for 2 separate experiments). (C) Tumor growth in WT mice injected subcutaneously with either B16F0 cells plus b3-null;Cas9 pericytes or B16F0 plus b3-null;AktKO pericytes at 1:8 ratio (n = 10 per group). (D) Western blot analysis using mock- and Ik-Ba-SR-transfected WT or b3-null pericyte lysates. HSC70, loading control. (E) Protein-profiler cytokine arrays. Quantitation of fold difference in cytokine expression between mock- and Ik-Ba-SR-transfected b3-null pericytes (n = 4 dots from 2 independent experiments). (F) Conditioned media from mock- and Ik-Ba-SR-transfected b3-null pericytes were applied to B16F0, B16F10, and LLC cells and cell survival measured by MTS assay. Bar charts, means ± SEM relative to tumor cells treated with conditioned media from mock-transfected b3-null pericytes (n = 3 experimental repeats).
Lentiviral Vector 32, 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
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Addgene inc plvuth
Figure 6. Deletion of Pericyte-b3-Integrin Enhances Tumor Cell Growth via Pericyte-pAkt-p65-Regulated Cytokine Production (A) Western blot analysis of b3-null pericytes with CRISPR-Cas9 deletion of <t>Akt1</t> (b3-null;AktKO PC) or HGFR (b3-null;HGFRKO PC) compared with b3-null;Cas9 controls. Hsc70, loading control. Blots representative of 3 separate experiments. Bar charts, average densitometric analysis relative to loading controls (n = 3 experimental repeats). (B) B16F0 cell survival after exposure to conditioned media from b3-null;AktKO, b3-null;HGFRKO, or b3-null;Cas9 pericytes (n = 16 technical replicates for 2 separate experiments). (C) Tumor growth in WT mice injected subcutaneously with either B16F0 cells plus b3-null;Cas9 pericytes or B16F0 plus b3-null;AktKO pericytes at 1:8 ratio (n = 10 per group). (D) Western blot analysis using mock- and Ik-Ba-SR-transfected WT or b3-null pericyte lysates. HSC70, loading control. (E) Protein-profiler cytokine arrays. Quantitation of fold difference in cytokine expression between mock- and Ik-Ba-SR-transfected b3-null pericytes (n = 4 dots from 2 independent experiments). (F) Conditioned media from mock- and Ik-Ba-SR-transfected b3-null pericytes were applied to B16F0, B16F10, and LLC cells and cell survival measured by MTS assay. Bar charts, means ± SEM relative to tumor cells treated with conditioned media from mock-transfected b3-null pericytes (n = 3 experimental repeats).
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Proteintech anti ttr
Figure 6. Deletion of Pericyte-b3-Integrin Enhances Tumor Cell Growth via Pericyte-pAkt-p65-Regulated Cytokine Production (A) Western blot analysis of b3-null pericytes with CRISPR-Cas9 deletion of <t>Akt1</t> (b3-null;AktKO PC) or HGFR (b3-null;HGFRKO PC) compared with b3-null;Cas9 controls. Hsc70, loading control. Blots representative of 3 separate experiments. Bar charts, average densitometric analysis relative to loading controls (n = 3 experimental repeats). (B) B16F0 cell survival after exposure to conditioned media from b3-null;AktKO, b3-null;HGFRKO, or b3-null;Cas9 pericytes (n = 16 technical replicates for 2 separate experiments). (C) Tumor growth in WT mice injected subcutaneously with either B16F0 cells plus b3-null;Cas9 pericytes or B16F0 plus b3-null;AktKO pericytes at 1:8 ratio (n = 10 per group). (D) Western blot analysis using mock- and Ik-Ba-SR-transfected WT or b3-null pericyte lysates. HSC70, loading control. (E) Protein-profiler cytokine arrays. Quantitation of fold difference in cytokine expression between mock- and Ik-Ba-SR-transfected b3-null pericytes (n = 4 dots from 2 independent experiments). (F) Conditioned media from mock- and Ik-Ba-SR-transfected b3-null pericytes were applied to B16F0, B16F10, and LLC cells and cell survival measured by MTS assay. Bar charts, means ± SEM relative to tumor cells treated with conditioned media from mock-transfected b3-null pericytes (n = 3 experimental repeats).
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Image Search Results


Journal: STAR Protocols

Article Title: Streamlined high-throughput cloning protocol to generate arrayed mutant libraries

doi: 10.1016/j.xpro.2022.101930

Figure Lengend Snippet:

Article Snippet: Microplate Rack for Incubator Shaker , Eppendorf , Cat#TTR-221.

Techniques: Virus, Recombinant, Plasmid Preparation, Mutagenesis, Software, Sterility, Aerosol

Figure 6. Deletion of Pericyte-b3-Integrin Enhances Tumor Cell Growth via Pericyte-pAkt-p65-Regulated Cytokine Production (A) Western blot analysis of b3-null pericytes with CRISPR-Cas9 deletion of Akt1 (b3-null;AktKO PC) or HGFR (b3-null;HGFRKO PC) compared with b3-null;Cas9 controls. Hsc70, loading control. Blots representative of 3 separate experiments. Bar charts, average densitometric analysis relative to loading controls (n = 3 experimental repeats). (B) B16F0 cell survival after exposure to conditioned media from b3-null;AktKO, b3-null;HGFRKO, or b3-null;Cas9 pericytes (n = 16 technical replicates for 2 separate experiments). (C) Tumor growth in WT mice injected subcutaneously with either B16F0 cells plus b3-null;Cas9 pericytes or B16F0 plus b3-null;AktKO pericytes at 1:8 ratio (n = 10 per group). (D) Western blot analysis using mock- and Ik-Ba-SR-transfected WT or b3-null pericyte lysates. HSC70, loading control. (E) Protein-profiler cytokine arrays. Quantitation of fold difference in cytokine expression between mock- and Ik-Ba-SR-transfected b3-null pericytes (n = 4 dots from 2 independent experiments). (F) Conditioned media from mock- and Ik-Ba-SR-transfected b3-null pericytes were applied to B16F0, B16F10, and LLC cells and cell survival measured by MTS assay. Bar charts, means ± SEM relative to tumor cells treated with conditioned media from mock-transfected b3-null pericytes (n = 3 experimental repeats).

Journal: Cell

Article Title: Cancer Burden Is Controlled by Mural Cell-β3-Integrin Regulated Crosstalk with Tumor Cells.

doi: 10.1016/j.cell.2020.02.003

Figure Lengend Snippet: Figure 6. Deletion of Pericyte-b3-Integrin Enhances Tumor Cell Growth via Pericyte-pAkt-p65-Regulated Cytokine Production (A) Western blot analysis of b3-null pericytes with CRISPR-Cas9 deletion of Akt1 (b3-null;AktKO PC) or HGFR (b3-null;HGFRKO PC) compared with b3-null;Cas9 controls. Hsc70, loading control. Blots representative of 3 separate experiments. Bar charts, average densitometric analysis relative to loading controls (n = 3 experimental repeats). (B) B16F0 cell survival after exposure to conditioned media from b3-null;AktKO, b3-null;HGFRKO, or b3-null;Cas9 pericytes (n = 16 technical replicates for 2 separate experiments). (C) Tumor growth in WT mice injected subcutaneously with either B16F0 cells plus b3-null;Cas9 pericytes or B16F0 plus b3-null;AktKO pericytes at 1:8 ratio (n = 10 per group). (D) Western blot analysis using mock- and Ik-Ba-SR-transfected WT or b3-null pericyte lysates. HSC70, loading control. (E) Protein-profiler cytokine arrays. Quantitation of fold difference in cytokine expression between mock- and Ik-Ba-SR-transfected b3-null pericytes (n = 4 dots from 2 independent experiments). (F) Conditioned media from mock- and Ik-Ba-SR-transfected b3-null pericytes were applied to B16F0, B16F10, and LLC cells and cell survival measured by MTS assay. Bar charts, means ± SEM relative to tumor cells treated with conditioned media from mock-transfected b3-null pericytes (n = 3 experimental repeats).

Article Snippet: SgRNAs were designed using the CRISPOR algorithm (http://crispor.tefor.net). sgRNA sequences targeting TIMP-1 (Gene ID: 21857), CXCL1 (Gene ID: 14825), CCL2 (Gene ID: 20296), Akt1 (Gene ID: 11651), and HGFR (Met) (Gene ID: 17295) (see Table S1) were cloned into the pLenti-CRISPR–EGFP plasmid (Addgene-#75159) using BsmBI enzyme site.

Techniques: Western Blot, CRISPR, Control, Injection, Transfection, Quantitation Assay, Expressing, MTS Assay