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<t>SLIT3</t> deficiency predicted poor prognosis in patients with NSCLC. (a) Kaplan-Meier overall survival curve was shown according to high and low expression of SLIT1∼3 in NSCLC patients ( n = 513) derived from TCGA data. (b) mRNA expression of SLIT3 in 513 NSCLC tissues and 397 normal tissues derived from TCGA data. (c) Western blotting of SLIT3 in A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (d) Cell proliferation of A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (e) Cell migration of A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (f) Immunostaining of SLIT3 in NSCLC tissues from high-degree (HD, stage III–IV) and low-degree (LD, stage I–II) patients ( n = 20). The scale bar was 100 μm.
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SLIT3 deficiency stimulated <t>UBE2C</t> upregulation. (a) 230 NSCLC patients from the TCGA database were categorized into SLIT3-high and -low groups, and the top 20 differentially expressed genes between these groups were enriched. (b) the correlation analysis between SLIT3 and UBE3C was conducted in 230 NSCLC patients. (c) mRNA expression of UBE2C in 513 NSCLC tissues and 397 normal tissues derived from TCGA data. (d) Kaplan-Meier overall survival curve was shown according to high and low expression of UBE2C in NSCLC patients ( n = 513) derived from TCGA data. (e) Western blotting of UBE2C in A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (f) SLIT3 siRNAs treated A549/NCI-H1299 cells were treated with scramble and UBE2C siRNAs. Then the expression of UBE2C were examined by western blotting. (g) Cell proliferation of A549/NCI-H1299 cells in (f). (h) Cell migration of A549/NCI-H1299 cells in (f). (i) Immunostaining of UBE2C in NSCLC tissues from high-degree (HD, stage III–IV) and low-degree (LD, stage I–II) patients ( n = 20). The scale bar was 100 μm.
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SLIT3 deficiency stimulated <t>UBE2C</t> upregulation. (a) 230 NSCLC patients from the TCGA database were categorized into SLIT3-high and -low groups, and the top 20 differentially expressed genes between these groups were enriched. (b) the correlation analysis between SLIT3 and UBE3C was conducted in 230 NSCLC patients. (c) mRNA expression of UBE2C in 513 NSCLC tissues and 397 normal tissues derived from TCGA data. (d) Kaplan-Meier overall survival curve was shown according to high and low expression of UBE2C in NSCLC patients ( n = 513) derived from TCGA data. (e) Western blotting of UBE2C in A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (f) SLIT3 siRNAs treated A549/NCI-H1299 cells were treated with scramble and UBE2C siRNAs. Then the expression of UBE2C were examined by western blotting. (g) Cell proliferation of A549/NCI-H1299 cells in (f). (h) Cell migration of A549/NCI-H1299 cells in (f). (i) Immunostaining of UBE2C in NSCLC tissues from high-degree (HD, stage III–IV) and low-degree (LD, stage I–II) patients ( n = 20). The scale bar was 100 μm.
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SLIT3 deficiency predicted poor prognosis in patients with NSCLC. (a) Kaplan-Meier overall survival curve was shown according to high and low expression of SLIT1∼3 in NSCLC patients ( n = 513) derived from TCGA data. (b) mRNA expression of SLIT3 in 513 NSCLC tissues and 397 normal tissues derived from TCGA data. (c) Western blotting of SLIT3 in A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (d) Cell proliferation of A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (e) Cell migration of A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (f) Immunostaining of SLIT3 in NSCLC tissues from high-degree (HD, stage III–IV) and low-degree (LD, stage I–II) patients ( n = 20). The scale bar was 100 μm.

Journal: Open Life Sciences

Article Title: SLIT3 deficiency promotes non-small cell lung cancer progression by modulating UBE2C/WNT signaling

doi: 10.1515/biol-2022-0956

Figure Lengend Snippet: SLIT3 deficiency predicted poor prognosis in patients with NSCLC. (a) Kaplan-Meier overall survival curve was shown according to high and low expression of SLIT1∼3 in NSCLC patients ( n = 513) derived from TCGA data. (b) mRNA expression of SLIT3 in 513 NSCLC tissues and 397 normal tissues derived from TCGA data. (c) Western blotting of SLIT3 in A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (d) Cell proliferation of A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (e) Cell migration of A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (f) Immunostaining of SLIT3 in NSCLC tissues from high-degree (HD, stage III–IV) and low-degree (LD, stage I–II) patients ( n = 20). The scale bar was 100 μm.

Article Snippet: To silence the expression of SLIT3 or UBE2C, cells were transfected with 50 nM of si-SLIT3 or si-UBE2C by using lipofectamine 3000 (Thermo Fisher, USA) for 48 h. The specific SLIT3 siRNA sequences were as follows: siRNA#1: 5′-GCAAAGAGCCGGUAGAUAUTT-3′ and siRNA#2: 5′-GUACAAAGAGCCAGGAAUATT-3′, and the specific UBE2C siRNA sequences were as follows: siRNA#1: 5′-GAAGTACCTGCAAGAAACCTACTCA-3 and siRNA#2: 5′-CAGCAGGAGCTGATGACCCTCATG-3′ (Tsingke Co, China).

Techniques: Expressing, Derivative Assay, Western Blot, Migration, Immunostaining

SLIT3 deficiency stimulated UBE2C upregulation. (a) 230 NSCLC patients from the TCGA database were categorized into SLIT3-high and -low groups, and the top 20 differentially expressed genes between these groups were enriched. (b) the correlation analysis between SLIT3 and UBE3C was conducted in 230 NSCLC patients. (c) mRNA expression of UBE2C in 513 NSCLC tissues and 397 normal tissues derived from TCGA data. (d) Kaplan-Meier overall survival curve was shown according to high and low expression of UBE2C in NSCLC patients ( n = 513) derived from TCGA data. (e) Western blotting of UBE2C in A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (f) SLIT3 siRNAs treated A549/NCI-H1299 cells were treated with scramble and UBE2C siRNAs. Then the expression of UBE2C were examined by western blotting. (g) Cell proliferation of A549/NCI-H1299 cells in (f). (h) Cell migration of A549/NCI-H1299 cells in (f). (i) Immunostaining of UBE2C in NSCLC tissues from high-degree (HD, stage III–IV) and low-degree (LD, stage I–II) patients ( n = 20). The scale bar was 100 μm.

Journal: Open Life Sciences

Article Title: SLIT3 deficiency promotes non-small cell lung cancer progression by modulating UBE2C/WNT signaling

doi: 10.1515/biol-2022-0956

Figure Lengend Snippet: SLIT3 deficiency stimulated UBE2C upregulation. (a) 230 NSCLC patients from the TCGA database were categorized into SLIT3-high and -low groups, and the top 20 differentially expressed genes between these groups were enriched. (b) the correlation analysis between SLIT3 and UBE3C was conducted in 230 NSCLC patients. (c) mRNA expression of UBE2C in 513 NSCLC tissues and 397 normal tissues derived from TCGA data. (d) Kaplan-Meier overall survival curve was shown according to high and low expression of UBE2C in NSCLC patients ( n = 513) derived from TCGA data. (e) Western blotting of UBE2C in A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (f) SLIT3 siRNAs treated A549/NCI-H1299 cells were treated with scramble and UBE2C siRNAs. Then the expression of UBE2C were examined by western blotting. (g) Cell proliferation of A549/NCI-H1299 cells in (f). (h) Cell migration of A549/NCI-H1299 cells in (f). (i) Immunostaining of UBE2C in NSCLC tissues from high-degree (HD, stage III–IV) and low-degree (LD, stage I–II) patients ( n = 20). The scale bar was 100 μm.

Article Snippet: To silence the expression of SLIT3 or UBE2C, cells were transfected with 50 nM of si-SLIT3 or si-UBE2C by using lipofectamine 3000 (Thermo Fisher, USA) for 48 h. The specific SLIT3 siRNA sequences were as follows: siRNA#1: 5′-GCAAAGAGCCGGUAGAUAUTT-3′ and siRNA#2: 5′-GUACAAAGAGCCAGGAAUATT-3′, and the specific UBE2C siRNA sequences were as follows: siRNA#1: 5′-GAAGTACCTGCAAGAAACCTACTCA-3 and siRNA#2: 5′-CAGCAGGAGCTGATGACCCTCATG-3′ (Tsingke Co, China).

Techniques: Expressing, Derivative Assay, Western Blot, Migration, Immunostaining

SLIT3/UBE2C axis modulated NSCLC progression through β-catenin/Wnt3A signaling. (a) the heatmap of Wnt1, Wnt2, and Wnt3A expression in A549 cells treated with scramble, UBE2C and SLIT3 siRNAs, respectively. (b) Western blotting of Wnt3A and β-catenin in A549/NCI-H1299 cells treated with scramble, UBE2C and SLIT3 siRNAs, respectively. (c) Immunostaining of Wnt3A in NSCLC tissues from high-degree (HD, stage III–IV) and low-degree (LD, stage I–II) patients ( n = 20). The scale bar was 100 μm. (d) Cell proliferation of SLIT3 siRNAs pre-treated A549/NCI-H1299 cells, treated with PBS or Wnt-C59 (20 nM). (e) Cell migration of SLIT3 siRNAs pre-treated A549/NCI-H1299 cells, treated with PBS or Wnt-C59 (20 nM).

Journal: Open Life Sciences

Article Title: SLIT3 deficiency promotes non-small cell lung cancer progression by modulating UBE2C/WNT signaling

doi: 10.1515/biol-2022-0956

Figure Lengend Snippet: SLIT3/UBE2C axis modulated NSCLC progression through β-catenin/Wnt3A signaling. (a) the heatmap of Wnt1, Wnt2, and Wnt3A expression in A549 cells treated with scramble, UBE2C and SLIT3 siRNAs, respectively. (b) Western blotting of Wnt3A and β-catenin in A549/NCI-H1299 cells treated with scramble, UBE2C and SLIT3 siRNAs, respectively. (c) Immunostaining of Wnt3A in NSCLC tissues from high-degree (HD, stage III–IV) and low-degree (LD, stage I–II) patients ( n = 20). The scale bar was 100 μm. (d) Cell proliferation of SLIT3 siRNAs pre-treated A549/NCI-H1299 cells, treated with PBS or Wnt-C59 (20 nM). (e) Cell migration of SLIT3 siRNAs pre-treated A549/NCI-H1299 cells, treated with PBS or Wnt-C59 (20 nM).

Article Snippet: To silence the expression of SLIT3 or UBE2C, cells were transfected with 50 nM of si-SLIT3 or si-UBE2C by using lipofectamine 3000 (Thermo Fisher, USA) for 48 h. The specific SLIT3 siRNA sequences were as follows: siRNA#1: 5′-GCAAAGAGCCGGUAGAUAUTT-3′ and siRNA#2: 5′-GUACAAAGAGCCAGGAAUATT-3′, and the specific UBE2C siRNA sequences were as follows: siRNA#1: 5′-GAAGTACCTGCAAGAAACCTACTCA-3 and siRNA#2: 5′-CAGCAGGAGCTGATGACCCTCATG-3′ (Tsingke Co, China).

Techniques: Expressing, Western Blot, Immunostaining, Migration

Inhibition of Wnt signaling improved the outcome of chemotherapy. (a) and (b) A549 and NCI-H1299 cells were pre-treated with scramble or SLIT3 siRNAs. Then cells were treated with Cis (1 μg/ml) or PTX (1 μg/ml) for 48 h. Cell apoptosis was examined. (c) and (d) A549 bearing mice ( n = 5 in each group) were treated with scramble or SLIT3 siRNAs via intratumor injection on days 10, 15, and 20. Tumor volume and survival information were recorded. (e) and (f) SLIT3 siRNAs treated tumor-bearing mice ( n = 5 in each group) were treated with PBS, PTX (10 mg/kg), Wnt-C59 (10 mg/kg), or a combination of them via tail vein injection on days 10, 15, and 20. Tumor volume and survival information were recorded.

Journal: Open Life Sciences

Article Title: SLIT3 deficiency promotes non-small cell lung cancer progression by modulating UBE2C/WNT signaling

doi: 10.1515/biol-2022-0956

Figure Lengend Snippet: Inhibition of Wnt signaling improved the outcome of chemotherapy. (a) and (b) A549 and NCI-H1299 cells were pre-treated with scramble or SLIT3 siRNAs. Then cells were treated with Cis (1 μg/ml) or PTX (1 μg/ml) for 48 h. Cell apoptosis was examined. (c) and (d) A549 bearing mice ( n = 5 in each group) were treated with scramble or SLIT3 siRNAs via intratumor injection on days 10, 15, and 20. Tumor volume and survival information were recorded. (e) and (f) SLIT3 siRNAs treated tumor-bearing mice ( n = 5 in each group) were treated with PBS, PTX (10 mg/kg), Wnt-C59 (10 mg/kg), or a combination of them via tail vein injection on days 10, 15, and 20. Tumor volume and survival information were recorded.

Article Snippet: To silence the expression of SLIT3 or UBE2C, cells were transfected with 50 nM of si-SLIT3 or si-UBE2C by using lipofectamine 3000 (Thermo Fisher, USA) for 48 h. The specific SLIT3 siRNA sequences were as follows: siRNA#1: 5′-GCAAAGAGCCGGUAGAUAUTT-3′ and siRNA#2: 5′-GUACAAAGAGCCAGGAAUATT-3′, and the specific UBE2C siRNA sequences were as follows: siRNA#1: 5′-GAAGTACCTGCAAGAAACCTACTCA-3 and siRNA#2: 5′-CAGCAGGAGCTGATGACCCTCATG-3′ (Tsingke Co, China).

Techniques: Inhibition, Injection

SLIT3 deficiency stimulated UBE2C upregulation. (a) 230 NSCLC patients from the TCGA database were categorized into SLIT3-high and -low groups, and the top 20 differentially expressed genes between these groups were enriched. (b) the correlation analysis between SLIT3 and UBE3C was conducted in 230 NSCLC patients. (c) mRNA expression of UBE2C in 513 NSCLC tissues and 397 normal tissues derived from TCGA data. (d) Kaplan-Meier overall survival curve was shown according to high and low expression of UBE2C in NSCLC patients ( n = 513) derived from TCGA data. (e) Western blotting of UBE2C in A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (f) SLIT3 siRNAs treated A549/NCI-H1299 cells were treated with scramble and UBE2C siRNAs. Then the expression of UBE2C were examined by western blotting. (g) Cell proliferation of A549/NCI-H1299 cells in (f). (h) Cell migration of A549/NCI-H1299 cells in (f). (i) Immunostaining of UBE2C in NSCLC tissues from high-degree (HD, stage III–IV) and low-degree (LD, stage I–II) patients ( n = 20). The scale bar was 100 μm.

Journal: Open Life Sciences

Article Title: SLIT3 deficiency promotes non-small cell lung cancer progression by modulating UBE2C/WNT signaling

doi: 10.1515/biol-2022-0956

Figure Lengend Snippet: SLIT3 deficiency stimulated UBE2C upregulation. (a) 230 NSCLC patients from the TCGA database were categorized into SLIT3-high and -low groups, and the top 20 differentially expressed genes between these groups were enriched. (b) the correlation analysis between SLIT3 and UBE3C was conducted in 230 NSCLC patients. (c) mRNA expression of UBE2C in 513 NSCLC tissues and 397 normal tissues derived from TCGA data. (d) Kaplan-Meier overall survival curve was shown according to high and low expression of UBE2C in NSCLC patients ( n = 513) derived from TCGA data. (e) Western blotting of UBE2C in A549/NCI-H1299 cells treated with scramble and SLIT3 siRNAs. (f) SLIT3 siRNAs treated A549/NCI-H1299 cells were treated with scramble and UBE2C siRNAs. Then the expression of UBE2C were examined by western blotting. (g) Cell proliferation of A549/NCI-H1299 cells in (f). (h) Cell migration of A549/NCI-H1299 cells in (f). (i) Immunostaining of UBE2C in NSCLC tissues from high-degree (HD, stage III–IV) and low-degree (LD, stage I–II) patients ( n = 20). The scale bar was 100 μm.

Article Snippet: To silence the expression of SLIT3 or UBE2C, cells were transfected with 50 nM of si-SLIT3 or si-UBE2C by using lipofectamine 3000 (Thermo Fisher, USA) for 48 h. The specific SLIT3 siRNA sequences were as follows: siRNA#1: 5′-GCAAAGAGCCGGUAGAUAUTT-3′ and siRNA#2: 5′-GUACAAAGAGCCAGGAAUATT-3′, and the specific UBE2C siRNA sequences were as follows: siRNA#1: 5′-GAAGTACCTGCAAGAAACCTACTCA-3 and siRNA#2: 5′-CAGCAGGAGCTGATGACCCTCATG-3′ (Tsingke Co, China).

Techniques: Expressing, Derivative Assay, Western Blot, Migration, Immunostaining

SLIT3/UBE2C axis modulated NSCLC progression through β-catenin/Wnt3A signaling. (a) the heatmap of Wnt1, Wnt2, and Wnt3A expression in A549 cells treated with scramble, UBE2C and SLIT3 siRNAs, respectively. (b) Western blotting of Wnt3A and β-catenin in A549/NCI-H1299 cells treated with scramble, UBE2C and SLIT3 siRNAs, respectively. (c) Immunostaining of Wnt3A in NSCLC tissues from high-degree (HD, stage III–IV) and low-degree (LD, stage I–II) patients ( n = 20). The scale bar was 100 μm. (d) Cell proliferation of SLIT3 siRNAs pre-treated A549/NCI-H1299 cells, treated with PBS or Wnt-C59 (20 nM). (e) Cell migration of SLIT3 siRNAs pre-treated A549/NCI-H1299 cells, treated with PBS or Wnt-C59 (20 nM).

Journal: Open Life Sciences

Article Title: SLIT3 deficiency promotes non-small cell lung cancer progression by modulating UBE2C/WNT signaling

doi: 10.1515/biol-2022-0956

Figure Lengend Snippet: SLIT3/UBE2C axis modulated NSCLC progression through β-catenin/Wnt3A signaling. (a) the heatmap of Wnt1, Wnt2, and Wnt3A expression in A549 cells treated with scramble, UBE2C and SLIT3 siRNAs, respectively. (b) Western blotting of Wnt3A and β-catenin in A549/NCI-H1299 cells treated with scramble, UBE2C and SLIT3 siRNAs, respectively. (c) Immunostaining of Wnt3A in NSCLC tissues from high-degree (HD, stage III–IV) and low-degree (LD, stage I–II) patients ( n = 20). The scale bar was 100 μm. (d) Cell proliferation of SLIT3 siRNAs pre-treated A549/NCI-H1299 cells, treated with PBS or Wnt-C59 (20 nM). (e) Cell migration of SLIT3 siRNAs pre-treated A549/NCI-H1299 cells, treated with PBS or Wnt-C59 (20 nM).

Article Snippet: To silence the expression of SLIT3 or UBE2C, cells were transfected with 50 nM of si-SLIT3 or si-UBE2C by using lipofectamine 3000 (Thermo Fisher, USA) for 48 h. The specific SLIT3 siRNA sequences were as follows: siRNA#1: 5′-GCAAAGAGCCGGUAGAUAUTT-3′ and siRNA#2: 5′-GUACAAAGAGCCAGGAAUATT-3′, and the specific UBE2C siRNA sequences were as follows: siRNA#1: 5′-GAAGTACCTGCAAGAAACCTACTCA-3 and siRNA#2: 5′-CAGCAGGAGCTGATGACCCTCATG-3′ (Tsingke Co, China).

Techniques: Expressing, Western Blot, Immunostaining, Migration