erbb2 tyrosine kinase inhibitor  (Bacus Laboratories)

Journal: The Journal of allergy and clinical immunology

Article Title: Dysfunctional ErbB2, an EGF receptor family member, in asthmatic airway epithelial cells critically hinders repair after mechanical wounding

doi: 10.1016/j.jaci.2018.11.046

Figure Lengend Snippet: Subject demographics

Article Snippet: For the ErbB2 inhibitor experiments, cultured cells were treated with a selective tyrosine kinase inhibitor of ErbB2 (mubritinib, Selleck Chemicals, Houston, TX; 100 nM) or DMSO 16 hours prior to wounding until the time of harvest ( 29 ).

Techniques: In Vivo, In Vitro, Western Blot

Journal: The Journal of allergy and clinical immunology

Article Title: Dysfunctional ErbB2, an EGF receptor family member, in asthmatic airway epithelial cells critically hinders repair after mechanical wounding

doi: 10.1016/j.jaci.2018.11.046

Figure Lengend Snippet: Total and phosphorylated ErbB2 (tErbB2 and pErbB2) protein expression in freshly isolated HAECs was analyzed by western blotting (A-G) and immunofluorescent staining of cytospin samples (H-I). (A, D) There were no differences in tErbB2 protein expression across the groups. (B, E) pErbB2 protein and (C, F) the ratio of phosphorylated to total ErbB2 (p/t ErbB2) was lower in asthmatic HAECs, but without difference by severity. (G) Representative western blots for ErbB2 among freshly isolated HAECs. (H) Representative immunofluorescent pictures for pErbB2 among cytospin HAECs samples. Green: pErbB2, Blue: nucleus. The white bar represents 100 μm. (I) pErbB2 positive cells were less in asthmatic HAECs than in healthy HAECs. Each bar represents median with interquartile range. Statistical analysis was performed by Wilcoxon rank sum test. Symbols; ○: healthy control, ▲: mild-to-moderate asthma, ■: severe asthma. Abbreviations: HC, Healthy control; MMA, Mild-to-moderate asthma; SA, Severe asthma.

Article Snippet: For the ErbB2 inhibitor experiments, cultured cells were treated with a selective tyrosine kinase inhibitor of ErbB2 (mubritinib, Selleck Chemicals, Houston, TX; 100 nM) or DMSO 16 hours prior to wounding until the time of harvest ( 29 ).

Techniques: Expressing, Isolation, Western Blot, Staining, Control

Journal: The Journal of allergy and clinical immunology

Article Title: Dysfunctional ErbB2, an EGF receptor family member, in asthmatic airway epithelial cells critically hinders repair after mechanical wounding

doi: 10.1016/j.jaci.2018.11.046

Figure Lengend Snippet: (A) CCND1 mRNA expression. (B) CCND1protein expression. (C) Representative western blot for ErbB2 and CCND1 protein expression with or without wounding. (D) Representative immunofluorescent staining images for CCND1 at wound leading edge. Red: CCND1, Blue: nucleus. The white bar represents 300 μm. (E) CCND1 positive cells along wound leading edge were higher in healthy as compared to asthmatic HAECs. Each bar represents mean ± SEM. Statistical analysis was performed by Student’s t test. Symbols; ○: healthy control, ▲: mild-to-moderate asthma, ■: severe asthma.

Article Snippet: For the ErbB2 inhibitor experiments, cultured cells were treated with a selective tyrosine kinase inhibitor of ErbB2 (mubritinib, Selleck Chemicals, Houston, TX; 100 nM) or DMSO 16 hours prior to wounding until the time of harvest ( 29 ).

Techniques: Expressing, Western Blot, Staining, Control

Journal: The Journal of allergy and clinical immunology

Article Title: Dysfunctional ErbB2, an EGF receptor family member, in asthmatic airway epithelial cells critically hinders repair after mechanical wounding

doi: 10.1016/j.jaci.2018.11.046

Figure Lengend Snippet: (A) ERBB2 mRNA expression. (B) Total ErbB2 protein expression. (C) Phosphorylated ErbB2 protein expression. (D) Ratio of phosphorylated to total ErbB2 protein (P = 0.09 and 0.04 for difference in ratio between HC and asthma before and after wounding, respectively). (E) Representative immunofluorescent staining pictures and their intensity profile for phosphorylated ErbB2 and F-actin at wound leading edge. Red: phosphorylated ErbB2, Green: F-actin, Blue: nucleus. The white bar represents 100 μm. (F) Phosphorylated ErbB2 expression along leading edge of the wound was higher in healthy as compared to asthmatic HAECs. Each bar represents mean ± SEM. Statistical analysis was performed by Student’s t test. Symbols; ○: healthy control, ▲: mild-to-moderate asthma, ■: severe asthma.

Article Snippet: For the ErbB2 inhibitor experiments, cultured cells were treated with a selective tyrosine kinase inhibitor of ErbB2 (mubritinib, Selleck Chemicals, Houston, TX; 100 nM) or DMSO 16 hours prior to wounding until the time of harvest ( 29 ).

Techniques: Expressing, Staining, Control

Journal: The Journal of allergy and clinical immunology

Article Title: Dysfunctional ErbB2, an EGF receptor family member, in asthmatic airway epithelial cells critically hinders repair after mechanical wounding

doi: 10.1016/j.jaci.2018.11.046

Figure Lengend Snippet: (A) Mubritinib (100nM) treatment decreased wound closure after 7 hours of wounding with greater impact in healthy HAECs as compared to asthmatic HAECs (P = 0.04 for difference in decrease in wound closure between HCs and asthma). (B) CCND1 protein expression was inhibited by mubritinib treatment in healthy HAECs, but not in asthmatic HAECs (P = 0.04 for difference between HC and asthma). (C) Representative western blot for ErbB2 and CCND1 protein expression with or without mubritinib treatment. (D) Phosphorylation of ErbB2 along leading edge of the wound was impaired by mubritinib primarily in healthy HAECs. Representative immunofluorescent staining for phosphorylated ErbB2 and F-actin at leading edge of the wound. Red: phosphorylated ErbB2, Green: F-actin, Blue: nucleus. The white bar represents 100 μm. (E) Intensity ratio of phosphorylated ErbB2 to F-actin at wound leading edge was decreased by mubritinib in healthy HAECs, not in asthmatic HAECs. Each bar represents mean ± SEM. Statistical analysis was performed by Student’s t test.

Article Snippet: For the ErbB2 inhibitor experiments, cultured cells were treated with a selective tyrosine kinase inhibitor of ErbB2 (mubritinib, Selleck Chemicals, Houston, TX; 100 nM) or DMSO 16 hours prior to wounding until the time of harvest ( 29 ).

Techniques: Expressing, Western Blot, Phospho-proteomics, Staining