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9 3f10  (ATCC)
90
ATCC 9 3f10
9 3f10, supplied by ATCC, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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93
DSMZ streptomyces sp
Streptomyces Sp, supplied by DSMZ, 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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92
Addgene inc pcdna5 frt to sbp tnrc6a
Pcdna5 Frt To Sbp Tnrc6a, 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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86
Santa Cruz Biotechnology netrin 1 shrna plasmid
Netrin 1 Shrna Plasmid, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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92
Santa Cruz Biotechnology netrin 1 sirnas
<t>Netrin-1</t> is overexpressed in metastatic breast tumors. (A) Expression profile of netrin-1 and UNC5H2 examined with quantitative real-time RT-PCR. QRT-PCR was performed by using total RNA extracted from 51 tumor biopsies. They were obtained from patients with tumors localized to the breast (N0), with only axillary node involvement (N + M0), and with distant metastases at diagnosis (M+). Specific human netrin-1 primers (35) and primers corresponding to the human HMBS gene (hydroxymethylbilane synthase) were used. HMBS was used as a reference here because it shows a weak variability at the mRNA level between normal and breast tumoral tissues, as described (36). Another reference TBP was also used with similar results (data not shown). Netrin-1 expression is given as the ratio between netrin-1 expression in each sample and the average of netrin-1 expression in the N0 samples, shown by a horizontal bar. UNC5H2 expression is also given as the ratio between UNC5H2 expression in each sample and the average of UNC5H2 level in N0 samples. Nonparametric statistical significance tests (Mann–Whitney) were used; the P values for netrin-1 are indicated. The average of netrin-1 and UNC5H2 expression is indicated for each tumor stage (upper left). (B) Representative netrin-1 immunohistochemistries on sections from two human tumors presented in A. Enlargement is indicated. (C) Comparison between netrin-1 or VEGFR expression and CXCR4 expression in seven pairs of frozen human invaded lymph nodes/primary tumors. Ratio between the mRNA level measured by QRT-PCR in invaded lymph node to the one detected in the primary tumor is shown for each pair and is represented as the ratio of netrin-1 (VEGFR) to CXCR4. (D) cDNA amplification from 67NR or 4T1 cells loaded in an agarose gel. (E) Immunostaining on 4T1 cell line using netrin-1 antibody. A control without the primary antibody is indicated. Nuclei were visualized with Hoescht staining.
Netrin 1 Sirnas, supplied by Santa Cruz Biotechnology, 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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Image Search Results


Netrin-1 is overexpressed in metastatic breast tumors. (A) Expression profile of netrin-1 and UNC5H2 examined with quantitative real-time RT-PCR. QRT-PCR was performed by using total RNA extracted from 51 tumor biopsies. They were obtained from patients with tumors localized to the breast (N0), with only axillary node involvement (N + M0), and with distant metastases at diagnosis (M+). Specific human netrin-1 primers (35) and primers corresponding to the human HMBS gene (hydroxymethylbilane synthase) were used. HMBS was used as a reference here because it shows a weak variability at the mRNA level between normal and breast tumoral tissues, as described (36). Another reference TBP was also used with similar results (data not shown). Netrin-1 expression is given as the ratio between netrin-1 expression in each sample and the average of netrin-1 expression in the N0 samples, shown by a horizontal bar. UNC5H2 expression is also given as the ratio between UNC5H2 expression in each sample and the average of UNC5H2 level in N0 samples. Nonparametric statistical significance tests (Mann–Whitney) were used; the P values for netrin-1 are indicated. The average of netrin-1 and UNC5H2 expression is indicated for each tumor stage (upper left). (B) Representative netrin-1 immunohistochemistries on sections from two human tumors presented in A. Enlargement is indicated. (C) Comparison between netrin-1 or VEGFR expression and CXCR4 expression in seven pairs of frozen human invaded lymph nodes/primary tumors. Ratio between the mRNA level measured by QRT-PCR in invaded lymph node to the one detected in the primary tumor is shown for each pair and is represented as the ratio of netrin-1 (VEGFR) to CXCR4. (D) cDNA amplification from 67NR or 4T1 cells loaded in an agarose gel. (E) Immunostaining on 4T1 cell line using netrin-1 antibody. A control without the primary antibody is indicated. Nuclei were visualized with Hoescht staining.

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Netrin-1 expression confers a selective advantage for tumor cell survival in metastatic breast cancer

doi: 10.1073/pnas.0709810105

Figure Lengend Snippet: Netrin-1 is overexpressed in metastatic breast tumors. (A) Expression profile of netrin-1 and UNC5H2 examined with quantitative real-time RT-PCR. QRT-PCR was performed by using total RNA extracted from 51 tumor biopsies. They were obtained from patients with tumors localized to the breast (N0), with only axillary node involvement (N + M0), and with distant metastases at diagnosis (M+). Specific human netrin-1 primers (35) and primers corresponding to the human HMBS gene (hydroxymethylbilane synthase) were used. HMBS was used as a reference here because it shows a weak variability at the mRNA level between normal and breast tumoral tissues, as described (36). Another reference TBP was also used with similar results (data not shown). Netrin-1 expression is given as the ratio between netrin-1 expression in each sample and the average of netrin-1 expression in the N0 samples, shown by a horizontal bar. UNC5H2 expression is also given as the ratio between UNC5H2 expression in each sample and the average of UNC5H2 level in N0 samples. Nonparametric statistical significance tests (Mann–Whitney) were used; the P values for netrin-1 are indicated. The average of netrin-1 and UNC5H2 expression is indicated for each tumor stage (upper left). (B) Representative netrin-1 immunohistochemistries on sections from two human tumors presented in A. Enlargement is indicated. (C) Comparison between netrin-1 or VEGFR expression and CXCR4 expression in seven pairs of frozen human invaded lymph nodes/primary tumors. Ratio between the mRNA level measured by QRT-PCR in invaded lymph node to the one detected in the primary tumor is shown for each pair and is represented as the ratio of netrin-1 (VEGFR) to CXCR4. (D) cDNA amplification from 67NR or 4T1 cells loaded in an agarose gel. (E) Immunostaining on 4T1 cell line using netrin-1 antibody. A control without the primary antibody is indicated. Nuclei were visualized with Hoescht staining.

Article Snippet: For cell culture use, scramble and netrin-1 siRNAs were designed by Santa Cruz Biotechnology as a pool of three target-specific 20- to 25-nt siRNAs.

Techniques: Expressing, Quantitative RT-PCR, MANN-WHITNEY, Amplification, Agarose Gel Electrophoresis, Immunostaining, Staining

Induction of metastatic cell death by inhibiting the netrin-1 autocrine loop. (A) Induction of 4T1 cell death by reducing netrin-1 level. The 4T1 cells were treated with scramble siRNA (control) or netrin-1 siRNA. Twenty-four hours later, the netrin-1 mRNA level was quantified by QRT-PCR (Left), and cell death was determined 1 day later by trypan blue-exclusion assay (Right). (B) Scheme representing netrin-1 and its receptors, DCC and UNC5H. (Inset) The extracellular part of the DCC receptor (DCC-EC-Fc) and the fifth fibronectin type III domain of DCC (DCC-5Fbn) used to induce cell death. (C–E) Quantitative analysis of cell death in 67NR and 4T1 cells treated with DCC-EC-Fc (C and D), with DCC-5Fbn (E) or with an unrelated soluble protein IL3R-EC-Fc. Cell death/survival was quantified by the trypan blue-exclusion assay (C), by caspase activity measurement by flow cytometry (D), or by MTT assay (E). In this latter test, netrin-1 was added in excess to reverse the effect of DCC-5Fbn. (F) Cell-death induction by DCC-5Fbn in SKBR7 and T47D cell lines but not in MDAMB231 cells. Cell survival was quantified by MTT assay (Right), and apoptosis was measured by caspase activity assay (Left). (G) Quantitative analysis of cell survival by MTT assay in the presence of either IL3R-Fc or DCC-5Fbn in 4T1 cells or 4T1 cells stably transfected with UNC5H2-IC D412N (DN UNC5H; i.e., the intracellular domain of UNCSH2 mutated in the caspase site acts as a dominant negative for UNC5H proapoptotic activity (SI Fig. 7D in SI Appendix). (Inset) Western blot using an anti-HA-UNC5H2 antibody was performed on the mock-transfected and DN UNC5H-expressing 4T1 cell lines. Standard deviations are indicated (n > 3). In C–G, Mann–Whitney tests were performed, and a P value is indicated.

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Netrin-1 expression confers a selective advantage for tumor cell survival in metastatic breast cancer

doi: 10.1073/pnas.0709810105

Figure Lengend Snippet: Induction of metastatic cell death by inhibiting the netrin-1 autocrine loop. (A) Induction of 4T1 cell death by reducing netrin-1 level. The 4T1 cells were treated with scramble siRNA (control) or netrin-1 siRNA. Twenty-four hours later, the netrin-1 mRNA level was quantified by QRT-PCR (Left), and cell death was determined 1 day later by trypan blue-exclusion assay (Right). (B) Scheme representing netrin-1 and its receptors, DCC and UNC5H. (Inset) The extracellular part of the DCC receptor (DCC-EC-Fc) and the fifth fibronectin type III domain of DCC (DCC-5Fbn) used to induce cell death. (C–E) Quantitative analysis of cell death in 67NR and 4T1 cells treated with DCC-EC-Fc (C and D), with DCC-5Fbn (E) or with an unrelated soluble protein IL3R-EC-Fc. Cell death/survival was quantified by the trypan blue-exclusion assay (C), by caspase activity measurement by flow cytometry (D), or by MTT assay (E). In this latter test, netrin-1 was added in excess to reverse the effect of DCC-5Fbn. (F) Cell-death induction by DCC-5Fbn in SKBR7 and T47D cell lines but not in MDAMB231 cells. Cell survival was quantified by MTT assay (Right), and apoptosis was measured by caspase activity assay (Left). (G) Quantitative analysis of cell survival by MTT assay in the presence of either IL3R-Fc or DCC-5Fbn in 4T1 cells or 4T1 cells stably transfected with UNC5H2-IC D412N (DN UNC5H; i.e., the intracellular domain of UNCSH2 mutated in the caspase site acts as a dominant negative for UNC5H proapoptotic activity (SI Fig. 7D in SI Appendix). (Inset) Western blot using an anti-HA-UNC5H2 antibody was performed on the mock-transfected and DN UNC5H-expressing 4T1 cell lines. Standard deviations are indicated (n > 3). In C–G, Mann–Whitney tests were performed, and a P value is indicated.

Article Snippet: For cell culture use, scramble and netrin-1 siRNAs were designed by Santa Cruz Biotechnology as a pool of three target-specific 20- to 25-nt siRNAs.

Techniques: Quantitative RT-PCR, Trypan Blue Exclusion Assay, Activity Assay, Flow Cytometry, MTT Assay, Caspase Activity Assay, Stable Transfection, Transfection, Dominant Negative Mutation, Western Blot, Expressing, MANN-WHITNEY

Inhibition of metastasis formation in mice by disruption of the netrin-1 autocrine loop. The 4T1-luc cells were i.v. injected into BALB/c mice at day 0. (A–C) Scramble siRNA or netrin-1 siRNA was i.v. injected daily, and after 9 (A and B) or 14 (C) days, lung metastasis was studied by luminescence recording (A and B) or by examination of lungs under a microscope (C). (A) A representative image of luminescence recording of scramble siRNA- (Left) or netrin-1 siRNA- (Right) treated mice. (B) Quantification of the luminescent signal measured by the NightOwLB II system. A luminescent signal for each group (control vs. netrin-1 siRNA) is given as a mean of the different ratios between summed luminescent signal in the lung area (photons/sec) and a background area defined in the animal body. Number of mice is indicated (n = 14). (C) Number of lung metastatic nodules in individual mice were counted after day 14 under a dissection microscope in three treated populations [scramble (control), first set of siRNA also used in A and B, and second set of siRNA]. (D–F) As in A–C except than PBS or DCC-5Fbn were injected once daily i.v. or once i.p. starting at day 0. After 13 days, metastasis development was studied by luminescence recording (D and E) or by examination of lungs under a microscope (F). (F) A representative macroscopic photograph of lungs from PBS- (Left) or DCC-5Fbn- (Right) treated mice (Lower). Total number of lung metastatic nodules in individual mice were counted in the two treated populations (+PBS, +DCC-5Fbn) (Upper). (G) As in F except that GST-DCC-5Fbn (instead of Flag-DCC-5Fbn) or an unrelated protein GST-FaDD was injected daily. (H) Dose effect of DCC-5Fbn. As in F except that different concentrations of DCC-5Fbn were injected daily; n = 21. (I) The 4T1 cells (1 × 103) were i.v. injected at day 0. At day 7 (at this day from one to eight micrometastasis are visible, data not shown), the mice were treated daily with PBS or DCC-5Fbn, and enumeration of metastasis was performed at day 11. (J) The 4T1 cells (5 × 105) were i.v. injected at day 0. At day 8, when at least 90 macrometastasis are visible, the mice were treated daily with PBS or DCC-5Fbn, and enumeration of metastasis was performed from day 8 to day 18 for PBS-treated mice, as we were required ethically to euthanize them, to day 20 for DCC-5Fbn-treated mice (each point represents three mice). (K) Overall survival of mice, which were i.v. injected with 106 4T1 cells, treated daily with PBS (black) or DCC-5Fbn (gray). P values were calculated by using the Kaplan–Meier test. Standard deviations are indicated. In B, C, E, G, and I, Mann–Whitney tests were performed, and a P value is indicated.

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Netrin-1 expression confers a selective advantage for tumor cell survival in metastatic breast cancer

doi: 10.1073/pnas.0709810105

Figure Lengend Snippet: Inhibition of metastasis formation in mice by disruption of the netrin-1 autocrine loop. The 4T1-luc cells were i.v. injected into BALB/c mice at day 0. (A–C) Scramble siRNA or netrin-1 siRNA was i.v. injected daily, and after 9 (A and B) or 14 (C) days, lung metastasis was studied by luminescence recording (A and B) or by examination of lungs under a microscope (C). (A) A representative image of luminescence recording of scramble siRNA- (Left) or netrin-1 siRNA- (Right) treated mice. (B) Quantification of the luminescent signal measured by the NightOwLB II system. A luminescent signal for each group (control vs. netrin-1 siRNA) is given as a mean of the different ratios between summed luminescent signal in the lung area (photons/sec) and a background area defined in the animal body. Number of mice is indicated (n = 14). (C) Number of lung metastatic nodules in individual mice were counted after day 14 under a dissection microscope in three treated populations [scramble (control), first set of siRNA also used in A and B, and second set of siRNA]. (D–F) As in A–C except than PBS or DCC-5Fbn were injected once daily i.v. or once i.p. starting at day 0. After 13 days, metastasis development was studied by luminescence recording (D and E) or by examination of lungs under a microscope (F). (F) A representative macroscopic photograph of lungs from PBS- (Left) or DCC-5Fbn- (Right) treated mice (Lower). Total number of lung metastatic nodules in individual mice were counted in the two treated populations (+PBS, +DCC-5Fbn) (Upper). (G) As in F except that GST-DCC-5Fbn (instead of Flag-DCC-5Fbn) or an unrelated protein GST-FaDD was injected daily. (H) Dose effect of DCC-5Fbn. As in F except that different concentrations of DCC-5Fbn were injected daily; n = 21. (I) The 4T1 cells (1 × 103) were i.v. injected at day 0. At day 7 (at this day from one to eight micrometastasis are visible, data not shown), the mice were treated daily with PBS or DCC-5Fbn, and enumeration of metastasis was performed at day 11. (J) The 4T1 cells (5 × 105) were i.v. injected at day 0. At day 8, when at least 90 macrometastasis are visible, the mice were treated daily with PBS or DCC-5Fbn, and enumeration of metastasis was performed from day 8 to day 18 for PBS-treated mice, as we were required ethically to euthanize them, to day 20 for DCC-5Fbn-treated mice (each point represents three mice). (K) Overall survival of mice, which were i.v. injected with 106 4T1 cells, treated daily with PBS (black) or DCC-5Fbn (gray). P values were calculated by using the Kaplan–Meier test. Standard deviations are indicated. In B, C, E, G, and I, Mann–Whitney tests were performed, and a P value is indicated.

Article Snippet: For cell culture use, scramble and netrin-1 siRNAs were designed by Santa Cruz Biotechnology as a pool of three target-specific 20- to 25-nt siRNAs.

Techniques: Inhibition, Injection, Microscopy, Dissection, MANN-WHITNEY

Disruption of the netrin-1 autocrine loop inhibits spontaneous lung metastasis from a xenografted human breast tumor. (A) Antitumor effect of DCC-5Fbn in nude mice xenografted with a fresh human breast tumor. A human breast tumor showing a high netrin-1 level was xenografted into nude mice, and when the tumor reached a palpable size, the mice were i.v. treated daily with PBS or DCC-5Fbn. Metastases were analyzed and enumerated by a pathologist. (B) Comparative low-power magnifications of lung metastases in control (a) and DCC-5Fbn-treated (b) animals; tumor cells, characterized by large size, abundant cytoplasm, and irregular nucleus, are arranged in small masses of variable diameter, scattered within the lung parenchyma. (c and d) Higher magnifications of lung metastases in a DCC-5Fbn-treated animal. Metastatic cells, revealed by an anti-human cytokeratin 7 antibody, form a small mass surrounded by a dense inflammatory reaction (c); at an advanced stage, tumor cells are dissociated by infiltrative inflammatory cells (d). (a and b) H&E staining, original magnifications ×200. (c and d) Immunoperoxidase, followed by nuclear counterstaining with Mayer's hematoxylin, original magnification ×400.

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Netrin-1 expression confers a selective advantage for tumor cell survival in metastatic breast cancer

doi: 10.1073/pnas.0709810105

Figure Lengend Snippet: Disruption of the netrin-1 autocrine loop inhibits spontaneous lung metastasis from a xenografted human breast tumor. (A) Antitumor effect of DCC-5Fbn in nude mice xenografted with a fresh human breast tumor. A human breast tumor showing a high netrin-1 level was xenografted into nude mice, and when the tumor reached a palpable size, the mice were i.v. treated daily with PBS or DCC-5Fbn. Metastases were analyzed and enumerated by a pathologist. (B) Comparative low-power magnifications of lung metastases in control (a) and DCC-5Fbn-treated (b) animals; tumor cells, characterized by large size, abundant cytoplasm, and irregular nucleus, are arranged in small masses of variable diameter, scattered within the lung parenchyma. (c and d) Higher magnifications of lung metastases in a DCC-5Fbn-treated animal. Metastatic cells, revealed by an anti-human cytokeratin 7 antibody, form a small mass surrounded by a dense inflammatory reaction (c); at an advanced stage, tumor cells are dissociated by infiltrative inflammatory cells (d). (a and b) H&E staining, original magnifications ×200. (c and d) Immunoperoxidase, followed by nuclear counterstaining with Mayer's hematoxylin, original magnification ×400.

Article Snippet: For cell culture use, scramble and netrin-1 siRNAs were designed by Santa Cruz Biotechnology as a pool of three target-specific 20- to 25-nt siRNAs.

Techniques: Staining