primary antibodies against cul4a  (Proteintech)

Journal: Oncology reports

Article Title: Effect of CUL4A on the metastatic potential of lung adenocarcinoma to the bone.

doi: 10.3892/or.2019.7448

Figure Lengend Snippet: Figure 1. Expression levels of CUL4A in lung adenocarcinoma cells. CUL4A expression was analyzed using (A) reverse transcription‑quantitative PCR and (B) western blotting in A549, H1299 and H460 cells. P<0.01 vs. H460. (C) mRNA and (D) protein expression levels of CUL4A in cells transfected with a CUL4A overexpression vector. P<0.001 vs. respective NC. (E) mRNA and (F) protein expression levels of CUL4A in cells transfected with the lentiviral knockdown vectors. P<0.001 vs. respective NC. CUL4A, cullin4A; NC, negative control; sh, short hairpin.

Article Snippet: Resolved proteins were transferred to PVDF membranes, and the membranes were blocked in 5% fat-free milk for 1 h at room temperature and incubated overnight at 4 ̊C with specific primary antibodies against CUL4A (dilution 1:500; cat. no. 14851-1-AP; ProteinTech Group, Inc.), β-actin (dilution 1:10,000; cat. no. 051M4892; Sigma-Aldrich; Merck KGaA), E-cadherin (dilution 1:1,000; cat. no. MABT26; Merck KGaA), Vimentin (dilution 1:1,000; cat. no. MABT26; Merck KGaA), ZEB1 (dilution 1:500; cat. no. 21544-1-AP; ProteinTech Group, Inc.).

Techniques: Expressing, Western Blot, Transfection, Over Expression, Plasmid Preparation, Knockdown, Negative Control

Journal: Oncology reports

Article Title: Effect of CUL4A on the metastatic potential of lung adenocarcinoma to the bone.

doi: 10.3892/or.2019.7448

Figure Lengend Snippet: Figure 2. Effect of CUL4A on the proliferation of lung adenocarcinoma cells. (A) Proliferation of A549‑NC and A549‑CUL4A cells determined using an MTT assay. (B) Colony formation assays of A549‑NC and A549‑CUL4A cells. (C) Proliferation of H1299‑NC and H1299‑CUL4A cells determined using an MTT assay. (D) Colony formation assays of H1299‑NC and H1299‑CUL4A cells. (E) MTT assay and (F) colony formation assay of H460‑NC and H460‑shCUL4A cells. *P<0.05, **P<0.01 vs. respective NC. CUL4A, cullin4A; OD, optical density; NC, negative control.

Article Snippet: Resolved proteins were transferred to PVDF membranes, and the membranes were blocked in 5% fat-free milk for 1 h at room temperature and incubated overnight at 4 ̊C with specific primary antibodies against CUL4A (dilution 1:500; cat. no. 14851-1-AP; ProteinTech Group, Inc.), β-actin (dilution 1:10,000; cat. no. 051M4892; Sigma-Aldrich; Merck KGaA), E-cadherin (dilution 1:1,000; cat. no. MABT26; Merck KGaA), Vimentin (dilution 1:1,000; cat. no. MABT26; Merck KGaA), ZEB1 (dilution 1:500; cat. no. 21544-1-AP; ProteinTech Group, Inc.).

Techniques: MTT Assay, Colony Assay, Negative Control

Journal: Oncology reports

Article Title: Effect of CUL4A on the metastatic potential of lung adenocarcinoma to the bone.

doi: 10.3892/or.2019.7448

Figure Lengend Snippet: Figure 3. Effect of CUL4A on metastasis of lung adenocarcinoma cells. (A) Representative images (x4 magnification) of a wound‑healing assay. Representative images (x200) and statistical analysis of invasion of (B) A549‑NC and A549‑CUL4A, (C) H1299‑NC and H1299‑CUL4A, and (D) H460‑NC and H460‑shCUL4A cells in a Transwell invasion assay. Data are expressed as the mean ± standard deviation. CUL4A, cullin4A; NC, negative control; sh, short hairpin.

Article Snippet: Resolved proteins were transferred to PVDF membranes, and the membranes were blocked in 5% fat-free milk for 1 h at room temperature and incubated overnight at 4 ̊C with specific primary antibodies against CUL4A (dilution 1:500; cat. no. 14851-1-AP; ProteinTech Group, Inc.), β-actin (dilution 1:10,000; cat. no. 051M4892; Sigma-Aldrich; Merck KGaA), E-cadherin (dilution 1:1,000; cat. no. MABT26; Merck KGaA), Vimentin (dilution 1:1,000; cat. no. MABT26; Merck KGaA), ZEB1 (dilution 1:500; cat. no. 21544-1-AP; ProteinTech Group, Inc.).

Techniques: Wound Healing Assay, Transwell Invasion Assay, Standard Deviation, Negative Control

Journal: Oncology reports

Article Title: Effect of CUL4A on the metastatic potential of lung adenocarcinoma to the bone.

doi: 10.3892/or.2019.7448

Figure Lengend Snippet: Figure 5. CUL4A mediates ZEB1‑induced epithelial‑mesenchymal transi- tion. Expression of ZEB1 and mesenchymal markers were upregulated and expression of epithelial markers were decreased in A549‑CUL4A and H1299‑CUL4A cells. Expression of mesenchymal proteins were decreased and expression of epithelial proteins were increased in the H460‑shCUL4A cells compared with the H460‑NC cells. CUL4A, cullin4A; ZEB1, zinc finger E‑box binding homeobox 1; NC, negative control; sh, short hairpin.

Article Snippet: Resolved proteins were transferred to PVDF membranes, and the membranes were blocked in 5% fat-free milk for 1 h at room temperature and incubated overnight at 4 ̊C with specific primary antibodies against CUL4A (dilution 1:500; cat. no. 14851-1-AP; ProteinTech Group, Inc.), β-actin (dilution 1:10,000; cat. no. 051M4892; Sigma-Aldrich; Merck KGaA), E-cadherin (dilution 1:1,000; cat. no. MABT26; Merck KGaA), Vimentin (dilution 1:1,000; cat. no. MABT26; Merck KGaA), ZEB1 (dilution 1:500; cat. no. 21544-1-AP; ProteinTech Group, Inc.).

Techniques: Expressing, Binding Assay, Negative Control

Journal: Oncology reports

Article Title: Effect of CUL4A on the metastatic potential of lung adenocarcinoma to the bone.

doi: 10.3892/or.2019.7448

Figure Lengend Snippet: Figure 4. D‑luciferin biofluorescence and representative X‑ray images of bone metastasis in the mouse models. Control mice injected with A549‑NC cells. Lung tumor development was assessed in vivo using (A) luciferase imaging and (B and C) normal bone tissues were detected using X‑ray imaging. Experimental mice were injected with A549‑CUL4A cells and metastasis to the bone was assessed in vivo using (D) luciferase imaging and as the arrows indicate (E and F) osteolytic bone metastasis lesions in the tibia in the mouse were visualized using X‑ray imaging. NC, negative control; CUL4A, cullin4A.

Article Snippet: Resolved proteins were transferred to PVDF membranes, and the membranes were blocked in 5% fat-free milk for 1 h at room temperature and incubated overnight at 4 ̊C with specific primary antibodies against CUL4A (dilution 1:500; cat. no. 14851-1-AP; ProteinTech Group, Inc.), β-actin (dilution 1:10,000; cat. no. 051M4892; Sigma-Aldrich; Merck KGaA), E-cadherin (dilution 1:1,000; cat. no. MABT26; Merck KGaA), Vimentin (dilution 1:1,000; cat. no. MABT26; Merck KGaA), ZEB1 (dilution 1:500; cat. no. 21544-1-AP; ProteinTech Group, Inc.).

Techniques: Control, Injection, In Vivo, Luciferase, Imaging, Negative Control

Journal: PLoS ONE

Article Title: Recurrent Amplification at 13q34 Targets at CUL4A , IRS2 , and TFDP1 As an Independent Adverse Prognosticator in Intrahepatic Cholangiocarcinoma

doi: 10.1371/journal.pone.0145388

Figure Lengend Snippet: Description of western blot and immunohistochemical antibodies.

Article Snippet: Primary antibodies against CUL4A, IRS2, and TFDP1 were used and followed by the PicTureTM-Plus kit (ZYMED®; 2nd Generation Polymer Detection System, San Francisco, CA, USA) ( ).

Techniques: Western Blot, Immunohistochemical staining

Journal: PLoS ONE

Article Title: Recurrent Amplification at 13q34 Targets at CUL4A , IRS2 , and TFDP1 As an Independent Adverse Prognosticator in Intrahepatic Cholangiocarcinoma

doi: 10.1371/journal.pone.0145388

Figure Lengend Snippet: Copy number changes of target genes at 13q34.

Article Snippet: Primary antibodies against CUL4A, IRS2, and TFDP1 were used and followed by the PicTureTM-Plus kit (ZYMED®; 2nd Generation Polymer Detection System, San Francisco, CA, USA) ( ).

Techniques:

Journal: PLoS ONE

Article Title: Recurrent Amplification at 13q34 Targets at CUL4A , IRS2 , and TFDP1 As an Independent Adverse Prognosticator in Intrahepatic Cholangiocarcinoma

doi: 10.1371/journal.pone.0145388

Figure Lengend Snippet: Independent predictive factors of disease-free survival by multivariate analysis.

Article Snippet: Primary antibodies against CUL4A, IRS2, and TFDP1 were used and followed by the PicTureTM-Plus kit (ZYMED®; 2nd Generation Polymer Detection System, San Francisco, CA, USA) ( ).

Techniques:

Journal: PLoS ONE

Article Title: Recurrent Amplification at 13q34 Targets at CUL4A , IRS2 , and TFDP1 As an Independent Adverse Prognosticator in Intrahepatic Cholangiocarcinoma

doi: 10.1371/journal.pone.0145388

Figure Lengend Snippet: Positive nuclear staining was observed for CUL 4A and TFDP1, and cytoplasmic staining for IRS2 in tumor cells with amplification of target genes (a, e, i), compared with normal bile ductules in the adjacent portal tracts (b, f, j). Loss of expression for CUL 4A, IRS2, and TFDP1 (c, g, k) was observed in tumor cells containing a deletion of target genes, in contrast to their normal counterparts (d, h, l). Original magnification: × 200.

Article Snippet: Primary antibodies against CUL4A, IRS2, and TFDP1 were used and followed by the PicTureTM-Plus kit (ZYMED®; 2nd Generation Polymer Detection System, San Francisco, CA, USA) ( ).

Techniques: Staining, Amplification, Expressing