her3 sirna mixture  (Thermo Fisher)

Journal: Oncotarget

Article Title: Novel functional anti-HER3 monoclonal antibodies with potent anti-cancer effects on various human epithelial cancers

doi: 10.18632/oncotarget.27414

Figure Lengend Snippet: ( A ) Scheme for the sandwich ELISA (sELISA) with soluble HER3-GFP proteins for the selection of anti-HER3 mAbs. ( B ) An example of first screening of anti-HER3 mAbs using sELISA. ( C ) Second and third FCM screenings of anti-HER3 mAbs using HEK293 transfectants expressing GFP-fused HER family proteins. ( D ) Representative FCM histograms showing the reactivity of anti-HER3 mAb (Ab4) with human cancer cell lines.

Article Snippet: Subconfluent LS-174T cell were seeded in each well of 6-well plates (Corning Japan, Tokyo) at a destiny of 3 × 10 5 cells/well, and transfected with HER3 siRNA mixture (ERBB3HSS140802, 140813 and 176604 Stealth RNAi, each 10 pmol, Invitrogen) in 2 mL medium using Lipofectamine RNAiMAX (Invitrogen).

Techniques: Sandwich ELISA, Selection, Expressing

Journal: Oncotarget

Article Title: Novel functional anti-HER3 monoclonal antibodies with potent anti-cancer effects on various human epithelial cancers

doi: 10.18632/oncotarget.27414

Figure Lengend Snippet: ( A ) Specificity of anti-HER3 mAbs was determined by siRNA-mediated knockdown in LS-LM4 cells. Histograms were shown by black (without anti-HER3 mAbs) and red (with anti-HER3 mAbs) lines. ( B ) Specificity of anti-HER3 mAbs was determined by and CRIPR/Cas9-based knockout in SW1116 cells. Histograms were shown by red (without anti-HER3 mAbs) and blue (with anti-HER3 mAbs) lines. ( C ) Nearest germline variable region (VH and VL) segments of mAbs are shown. Usage of different germlines in CDR segments is highlighted with different colors. ( D ) Heat map from sequence homology (%) of CDR amino acids (identity) was determined using Pairwise Sequence Alignment ( https://www.ebi.ac.uk/Tools/psa/ ).

Article Snippet: Subconfluent LS-174T cell were seeded in each well of 6-well plates (Corning Japan, Tokyo) at a destiny of 3 × 10 5 cells/well, and transfected with HER3 siRNA mixture (ERBB3HSS140802, 140813 and 176604 Stealth RNAi, each 10 pmol, Invitrogen) in 2 mL medium using Lipofectamine RNAiMAX (Invitrogen).

Techniques: Knock-Out, Sequencing

Journal: Oncotarget

Article Title: Novel functional anti-HER3 monoclonal antibodies with potent anti-cancer effects on various human epithelial cancers

doi: 10.18632/oncotarget.27414

Figure Lengend Snippet: Reactivity of anti-HER3 mAb (Ab4) with human various normal, non-cancer and cancer cell lines was analyzed by FCM, and mAb binding was shown by ΔMFI.

Article Snippet: Subconfluent LS-174T cell were seeded in each well of 6-well plates (Corning Japan, Tokyo) at a destiny of 3 × 10 5 cells/well, and transfected with HER3 siRNA mixture (ERBB3HSS140802, 140813 and 176604 Stealth RNAi, each 10 pmol, Invitrogen) in 2 mL medium using Lipofectamine RNAiMAX (Invitrogen).

Techniques: Binding Assay

Journal: Oncotarget

Article Title: Novel functional anti-HER3 monoclonal antibodies with potent anti-cancer effects on various human epithelial cancers

doi: 10.18632/oncotarget.27414

Figure Lengend Snippet: ( A ) Scheme for the epitope analysis of anti-HER3 rat mAbs with HER3 transfectants. ( B ) 3D bar graph for the binding inhibition (%) by all combinations of anti-HER3 rat mAbs. ( C ) Scheme for the Patritumab binding in the presence of excess anti-HER3 rat mAbs. ( D ) Binding of Patritumab to transfectants in the presence of control rat IgG or anti-HER3 rat mAbs.

Article Snippet: Subconfluent LS-174T cell were seeded in each well of 6-well plates (Corning Japan, Tokyo) at a destiny of 3 × 10 5 cells/well, and transfected with HER3 siRNA mixture (ERBB3HSS140802, 140813 and 176604 Stealth RNAi, each 10 pmol, Invitrogen) in 2 mL medium using Lipofectamine RNAiMAX (Invitrogen).

Techniques: Binding Assay, Inhibition

Journal: Oncotarget

Article Title: Novel functional anti-HER3 monoclonal antibodies with potent anti-cancer effects on various human epithelial cancers

doi: 10.18632/oncotarget.27414

Figure Lengend Snippet: ( A ) Scheme for the internalization of HER3 by mAbs. Human cancer cells were treated with anti-HER3 mAbs at 37° C or 4° C for 1~1.5 h, and cell-surface HER3 proteins were analyzed by FCM. ( B ) Internalization of cell-surface HER3 proteins of LS-174T and T47D by anti-HER3 mAbs was shown as the internalization (%). ( C ) Scheme for the binding inhibition of mAbs to HEK293 cells expressing HER3-GFP in the presence of NRG1 (upper). Representative binding of anti-HER3 mAb in the absence (-) or presence (+) of NRG1(lower). ( D ) Histogram (left) for the binding of mAb to HEK293 cells expressing HER3-GFP in NRG1(+) or NRG1(-). Bar graph (right) shows binding inhibition of anti-HER3 mAbs with HEK293 cells expressing HER3-GFP in NRG (+).

Article Snippet: Subconfluent LS-174T cell were seeded in each well of 6-well plates (Corning Japan, Tokyo) at a destiny of 3 × 10 5 cells/well, and transfected with HER3 siRNA mixture (ERBB3HSS140802, 140813 and 176604 Stealth RNAi, each 10 pmol, Invitrogen) in 2 mL medium using Lipofectamine RNAiMAX (Invitrogen).

Techniques: Binding Assay, Inhibition, Expressing

Journal: Oncotarget

Article Title: Novel functional anti-HER3 monoclonal antibodies with potent anti-cancer effects on various human epithelial cancers

doi: 10.18632/oncotarget.27414

Figure Lengend Snippet: ( A ) Inhibition of NRG1-induced tyrosine phosphorylation of HER3 proteins by anti-HER3 mAbs in human cancer cells. ( B ) Effects of anti-HER3 mAbs on the cell growth of NRG1-treated BT474 cells. ( C ) Immunohistochemical staining of human colon carcinoma tissues with anti-HER3 mAb (Ab1). ( D ) Immunofluorescent staining of human colon cancer-derived CTOS with anti-HER3 mAbs (Ab1 and Ab3).

Article Snippet: Subconfluent LS-174T cell were seeded in each well of 6-well plates (Corning Japan, Tokyo) at a destiny of 3 × 10 5 cells/well, and transfected with HER3 siRNA mixture (ERBB3HSS140802, 140813 and 176604 Stealth RNAi, each 10 pmol, Invitrogen) in 2 mL medium using Lipofectamine RNAiMAX (Invitrogen).

Techniques: Inhibition, Immunohistochemical staining, Staining, Derivative Assay

Journal: Oncotarget

Article Title: Novel functional anti-HER3 monoclonal antibodies with potent anti-cancer effects on various human epithelial cancers

doi: 10.18632/oncotarget.27414

Figure Lengend Snippet: ( A ) PCA by the binding inhibition analyses of anti-HER3 mAbs. ( B ) PCA by the amino acid identity of CDR of anti-HER3 mAbs. ( C ) Correlation diagram about seven anti-HER3 mAbs between %CDR homology and binding inhibition (%). ( D ) Summary table showing various features of seven anti-HER3 mAbs. ( E ) In vitro ffects of anti-HER3 mAb (Ab4) or patritumab on the cell growth of MCF7 cells with or without Erlotinib. ( F ) Anti-tumor effects of anti-HER3 mAb (Ab4) or patritumab on BT474 human breast cancer cells were evaluated ( n = 4). Anti-tumor effects of anti-HER3 mAb (Ab4) on LS-174T ( G , n = 3) and LS-LM4 ( H , n = 8) human colon cancer cells were evaluated.

Article Snippet: Subconfluent LS-174T cell were seeded in each well of 6-well plates (Corning Japan, Tokyo) at a destiny of 3 × 10 5 cells/well, and transfected with HER3 siRNA mixture (ERBB3HSS140802, 140813 and 176604 Stealth RNAi, each 10 pmol, Invitrogen) in 2 mL medium using Lipofectamine RNAiMAX (Invitrogen).

Techniques: Binding Assay, Inhibition, In Vitro