Journal: Cancer Cell

Article Title: A living biobank of patient-derived ductal carcinoma in situ mouse-intraductal xenografts identifies risk factors for invasive progression

doi: 10.1016/j.ccell.2023.04.002

Figure Lengend Snippet:

Article Snippet: The ERBB2-GFP lentivirus functionality was tested by transfecting HEK-293T cells (ATCC) and performing a Western Blot with a HER2/ErbB2 and pHER2/ErbB2 antibody (Cell signaling) blocked in milk powder.

Techniques: Labeling, Recombinant, Electron Microscopy, Plasmid Preparation, Blocking Assay, Software

Journal: Breast Cancer Research : BCR

Article Title: Fatty acid synthase phosphorylation: a novel therapeutic target in HER2-overexpressing breast cancer cells

doi: 10.1186/bcr2777

Figure Lengend Snippet: Association of FASN and HER2 in breast cancer cells . SKBR3 (A) and BT474 (B) cells were treated for one hour with: 1) no agent (control); 2) 50 ng/mL HRG; 3) 1 μM lapatinib or 4) 50 ng/mL HRG plus 1 μM lapatinib. The immunoprecipitated FASN complexes were subjected to Western blotting for FASN, HER2 and PT66. (C) Whole-cell lysates from SKBR3 (left panel) and BT474 cells (right panel) were subjected to Western blotting for pHER2 (Y1248), pHER3 (Y1289), pAkt (S473), pErk1/2, HER2, HER3, Akt, Erk1/2, FASN and β-actin.

Article Snippet: The following antibodies were used for Western blot analysis: mouse monoclonal antibodies detecting tyrosine phosphorylation (PT66) and β-actin (Sigma-Aldrich); rabbit polyclonal antibodies detecting pHER2 (Tyr 1248), pHER3 (Tyr 1289), pAkt (S473), pErk1/2, Akt, Erk1/2 and HER3 (Cell Signaling Technology, Inc., Danvers, MA, USA); mouse monoclonal antibodies detecting FASN, HER2 and HER3 (Santa Cruz) and an Alexa Flour 680 goat anti-mouse immunoglobulin (IgG) antibody, Alexa Flour 680 goat anti-rabbit IgG antibody and Alexa Flour 680 donkey anti-goat IgG antibody (Invitrogen Corporation, Carlsbad, CA, USA).

Techniques: Immunoprecipitation, Western Blot

Journal: Breast Cancer Research : BCR

Article Title: Fatty acid synthase phosphorylation: a novel therapeutic target in HER2-overexpressing breast cancer cells

doi: 10.1186/bcr2777

Figure Lengend Snippet: FASN phosphorylation in breast cancer cells treated with C75 . SKBR3 (A) and BT474 (B) cells were pretreated with 10 μM C75 for five hours and then treated for one additional hour with 1) no agent (control); 2) 50 ng/mL HRG; 3) 10 μM C75; or 4) 50 ng/mL HRG plus 10 μM C75. The immunoprecipitated FASN complexes were assessed for FASN, HER2 and PT66 by Western blotting. (C) Whole-cell lysates from SKBR3 and BT474 cells were subjected to Western blotting for pHER2 (Y1248), pHER3 (Y1289), pAkt (S473), pErk1/2, HER2, HER3, Akt, Erk1/2, FASN and β-actin.

Article Snippet: The following antibodies were used for Western blot analysis: mouse monoclonal antibodies detecting tyrosine phosphorylation (PT66) and β-actin (Sigma-Aldrich); rabbit polyclonal antibodies detecting pHER2 (Tyr 1248), pHER3 (Tyr 1289), pAkt (S473), pErk1/2, Akt, Erk1/2 and HER3 (Cell Signaling Technology, Inc., Danvers, MA, USA); mouse monoclonal antibodies detecting FASN, HER2 and HER3 (Santa Cruz) and an Alexa Flour 680 goat anti-mouse immunoglobulin (IgG) antibody, Alexa Flour 680 goat anti-rabbit IgG antibody and Alexa Flour 680 donkey anti-goat IgG antibody (Invitrogen Corporation, Carlsbad, CA, USA).

Techniques: Immunoprecipitation, Western Blot