Journal: The Journal of Biological Chemistry

Article Title: Surfactant protein A down-regulates epidermal growth factor receptor by mechanisms different from those of surfactant protein D

doi: 10.1074/jbc.M117.800771

Figure Lengend Snippet: SP-A binds to EGFR in A549 cells, H441 cells, and CHOK1 cells stably expressing EGFR. A, whole-cell lysate of A549 cells was immunoprecipitated (IP) with anti-EGFR monoclonal antibody Ab-11 or control IgG (0. 8 μg) at 4 °C for 16 h. The samples and BSA (200 ng/lane) were subjected to SDS-PAGE and transferred onto PVDF membranes. The membranes were incubated with or without SP-A (1 μg/ml) for 16 h. The membranes not incubated with SP-A were subjected to Western blotting using an anti-human EGFR monoclonal antibody (WB: EGFR). The membranes incubated with SP-A were then treated with an anti-human SP-A polyclonal antibody (SP-A pAb) or monoclonal antibodies PE10 (SP-A mAb (PE10)) or PC6 (SP-A mAb (PC6)), which was followed by incubation with HRP-labeled anti-rabbit IgG or anti-mouse IgG (upper panel). As a negative control, the membranes in the absence of incubation with SP-A were also incubated with an anti-human SP-A polyclonal antibody (WB: SP-A pAb) or monoclonal antibodies PE10 (WB: SP-A mAb (PE10)) or PC6 (WB: SP-A mAb (PC6)), which was followed by incubation with HRP-labeled anti-rabbit IgG or anti-mouse IgG (lower panel). The data are representative of three independent experiments. B and C, experimental paradigm described in A was performed in H441 cells (B) and CHOK1 cells stably expressing EGFR (C) by using an anti-human SP-A polyclonal antibody, which was followed by incubation with HRP-labeled anti-rabbit IgG. The data are representative of three independent experiments.

Article Snippet: The H441 human lung adenocarcinoma cell line was obtained from ATCC and maintained in RPMI 1640 medium (Sigma) with 10% (v/v) FCS.

Techniques: Stable Transfection, Expressing, Immunoprecipitation, Control, SDS Page, Incubation, Western Blot, Bioprocessing, Labeling, Negative Control

Journal: Oncology Letters

Article Title: Antitumor activity of nimotuzumab in combination with cisplatin in lung cancer cell line A549 in vitro

doi: 10.3892/ol.2018.7923

Figure Lengend Snippet: Cyclin D1 expression in A549 cells in groups A-E following treatment for 48 h. Lanes: A, nimotuzumab; B, cisplatin; C, nimotuzumab followed by cisplatin; D, nimotuzumab and cisplatin simultaneously; and E, untreated control.

Article Snippet: Cell culture The A549 human lung adenocarcinoma epithelial cell line (supplied by the Central Laboratory of Qinhuangdao No. 1 People's Hospital) was cultured in Dulbecco's modified Eagle's medium (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) containing 10% fetal bovine serum (FBS; Zhejiang Tianhang Biotechnology Co., Ltd., Huzhou, China), 1 U/ml penicillin and 1 mg/ml streptomycin at 37°C, with 5% CO 2 and 95% humidity.

Techniques: Expressing, Control

Journal: Theranostics

Article Title: Prevention of radiotherapy-induced pro-tumorigenic microenvironment by SFK inhibitors

doi: 10.7150/thno.100970

Figure Lengend Snippet: SFK-targeted inhibitors suppress direct radiation-induced myofibroblast activation. ( A ) Comparison gene expression of SRC, YES1 and fibrosis markers in HLMVEC, 24 h after 10 Gy direct irradiation. The NCBI public dataset, GSE179810, was used. ( B ) SFKs expression and activation by direct irradiation in BEAS-2B cells. ( C ) Myofibroblast activation by direct irradiation in MRC5 cells. ( D ) Myofibroblast activation and its mechanism following direct irradiation in BEAS-2B cells. Total protein levels are shown in . ( E, F ) Reversal of direct radiation-induced myofibroblast activation by SFK-inhibitors. Total protein levels are shown in . ( G ) Schematic illustration representing these results. All western blot analyses in this study were repeated three times independently.

Article Snippet: To determine if myofibroblast activation is directly induced by irradiation, we examined the National Center for Biotechnology Information (NCBI) public datasets on the human lung endothelial cell line HLMVEC.

Techniques: Activation Assay, Comparison, Gene Expression, Irradiation, Expressing, Western Blot