cdna encoding human add3 bc062559) potb7 vector  (Transomic Technologies Inc)

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Adducins inhibit lung cancer cell migration through mechanisms involving regulation of cell-matrix adhesion and cadherin-11 expression

doi: 10.1016/j.bbamcr.2018.10.001

Figure Lengend Snippet: (A) A comparative analysis of ADD1 and ADD3 mRNA expression levels in normal (n=59) and tumor (n=457) tissues from patients with lung adenocarcinoma. Error bars indicate the median absolute deviations. (B) Immunoblotting analysis of adducins expression in epithelial and mesenchymal-type non-small cell lung cancer cells. (C) Immunofluorescence labeling of ADD1 and ADD3 in non-tumorigenic bronchial epithelial cells (16HBE14o), epithelial-type (H1573) and mesenchymal-type (H1299) lung cancer cells. Arrows point on plasma membrane localization of ADD1 and ADD3 in epithelial-type cells. Arrowheads indicate cytoplasmic and nuclear localization of ADD1 in mesenchymal-type lung cancer cells. Scale bar, 20μm.

Article Snippet: AS2.neomycin vector encoding FLAG-tagged wild-type ADD1 was obtained from Dr. Hong-Chen Chen, National Chung Hsing University, Taiwan [ 45 ], A cDNA encoding human ADD3 (clone {"type":"entrez-nucleotide","attrs":{"text":"BC062559","term_id":"38512197","term_text":"BC062559"}} BC062559 in the pOTB7 vector) was obtained from Transomic Technologies (Huntsville, AL) and the ADD3 insert was cloned into the pLKO.AS2.neo vector.

Techniques: Expressing, Western Blot, Immunofluorescence, Labeling

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Adducins inhibit lung cancer cell migration through mechanisms involving regulation of cell-matrix adhesion and cadherin-11 expression

doi: 10.1016/j.bbamcr.2018.10.001

Figure Lengend Snippet: ADD1 expression was down-regulated H1573 cells using CRISPR/Cas9-mediated gene editing. (A) Immunoblotting analysis shows co-depletion of ADD1 and ADD3 by 4 different ADD1 small guide (sg) RNAs. (B) Quantification of the planar migration of the control and ADDl-depleted H1573 cell monolayers at 24 h post-wounding. (C, D) Representative images and quantitative analysis of the DAPI-labeled control and ADD 1-deficient HI 573 cells after 16 h of transfilter migration in the Boyden chamber. (E, F) Representative images and quantitative analysis of the DAPI-labeled control and ADD 1-deficient H1573 cells after 24 h invasion into Matrigel. Data are presented as mean ± SE (n =3); **p < 0.005, as compared to the control sgRNA-transfected group.

Article Snippet: AS2.neomycin vector encoding FLAG-tagged wild-type ADD1 was obtained from Dr. Hong-Chen Chen, National Chung Hsing University, Taiwan [ 45 ], A cDNA encoding human ADD3 (clone {"type":"entrez-nucleotide","attrs":{"text":"BC062559","term_id":"38512197","term_text":"BC062559"}} BC062559 in the pOTB7 vector) was obtained from Transomic Technologies (Huntsville, AL) and the ADD3 insert was cloned into the pLKO.AS2.neo vector.

Techniques: Expressing, CRISPR, Western Blot, Migration, Labeling, Transfection

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Adducins inhibit lung cancer cell migration through mechanisms involving regulation of cell-matrix adhesion and cadherin-11 expression

doi: 10.1016/j.bbamcr.2018.10.001

Figure Lengend Snippet: ADD3 expression was down-regulated HI573 cells using CRISPR/Cas9-mediated gene editing. (A) Immunoblotting analysis shows co-depletion of ADD3 and ADD1 by 4 different ADD3 small guide (sg) RNAs. (B) Quantification of the planar migration of the control and ADD3-depleted HI573 cell monolayers at 24 h post-wounding. (C, D) Representative images and quantitative analysis of the DAPI-labeled control and ADD3-deficient HI 573 cells after 16 h of transfilter migration in the Boyden chamber. (E, F) Representative images and quantitative analysis of the DAPI-labeled control and ADD3-deficient HI573 cells after 24 h invasion into Matrigel. Data are presented as mean ± SE (n =3); *p < 0.05; **p < 0.005, as compared to the control sgRNA-transfected group.

Article Snippet: AS2.neomycin vector encoding FLAG-tagged wild-type ADD1 was obtained from Dr. Hong-Chen Chen, National Chung Hsing University, Taiwan [ 45 ], A cDNA encoding human ADD3 (clone {"type":"entrez-nucleotide","attrs":{"text":"BC062559","term_id":"38512197","term_text":"BC062559"}} BC062559 in the pOTB7 vector) was obtained from Transomic Technologies (Huntsville, AL) and the ADD3 insert was cloned into the pLKO.AS2.neo vector.

Techniques: Expressing, CRISPR, Western Blot, Migration, Labeling, Transfection

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Adducins inhibit lung cancer cell migration through mechanisms involving regulation of cell-matrix adhesion and cadherin-11 expression

doi: 10.1016/j.bbamcr.2018.10.001

Figure Lengend Snippet: FLAG-tagged ADD1 or ADD3 were stably expressed in H1299 lung cancer cells using a lentiviral expression vector. (A) Immunoblotting analysis shows the levels of ADD1 and ADD3 proteins in the generated cell lines. (B) Quantification of the planar migration of the control, ADD1 or ADD3-overexpressing H1299 cell monolayers after 12 h of wound healing. (C, D) Representative images and quantitative analysis of the DAPI-labeled control, ADD1, or ADD3-overexpressing H1299 cells after 12 h transfilter migration in the Boyden chamber. (E, F) Representative images and quantitative analysis of the DAPI-labeled control, ADD1, or ADD3-overexpressing H1299 cells after 24 h invasion into Matrigel. Data are presented as mean ± SE (n =3); *p < 0.05; **p < 0.005, as compared to the control group.

Article Snippet: AS2.neomycin vector encoding FLAG-tagged wild-type ADD1 was obtained from Dr. Hong-Chen Chen, National Chung Hsing University, Taiwan [ 45 ], A cDNA encoding human ADD3 (clone {"type":"entrez-nucleotide","attrs":{"text":"BC062559","term_id":"38512197","term_text":"BC062559"}} BC062559 in the pOTB7 vector) was obtained from Transomic Technologies (Huntsville, AL) and the ADD3 insert was cloned into the pLKO.AS2.neo vector.

Techniques: Stable Transfection, Expressing, Plasmid Preparation, Western Blot, Generated, Migration, Labeling

Journal: Biochimica et biophysica acta. Molecular cell research

Article Title: Adducins inhibit lung cancer cell migration through mechanisms involving regulation of cell-matrix adhesion and cadherin-11 expression

doi: 10.1016/j.bbamcr.2018.10.001

Figure Lengend Snippet: (A, B) Representative immunoblots showing expression of different cadherins and densitometric quantification of the cadherin-11 protein level in the control, ADD1, or ADD3-depleted H1573 cells. (C, D) Cadherin-11 was transiently depleted by siRNA in control and ADD 1-deficient H1573 cells. (C) Immunoblotting analysis shows the efficiency of cadherin-11 depletion. (D) Results of the transfilter migration assay of the control and ADD 1-deficient H1573 cells with and without cadherin-11 depletion. (E, F) Transfilter migration data for the control and ADD 1-deficient H1573 cells treated with either anti-cadherin-11 antibody or control IgG. Data are presented as mean ± SE (n =3); *p < 0.05; **p < 0.005.

Article Snippet: AS2.neomycin vector encoding FLAG-tagged wild-type ADD1 was obtained from Dr. Hong-Chen Chen, National Chung Hsing University, Taiwan [ 45 ], A cDNA encoding human ADD3 (clone {"type":"entrez-nucleotide","attrs":{"text":"BC062559","term_id":"38512197","term_text":"BC062559"}} BC062559 in the pOTB7 vector) was obtained from Transomic Technologies (Huntsville, AL) and the ADD3 insert was cloned into the pLKO.AS2.neo vector.

Techniques: Western Blot, Expressing, Migration