Journal: Journal of Leukocyte Biology

Article Title: HER2-specific synthetic antigen receptor T cell therapy synergizes with radiotherapy to provide improved antitumor efficacy in non-small cell lung cancer

doi: 10.1093/jleuko/qiag030

Figure Lengend Snippet: Radiation-mediated regulation of HER2 expression in NSCLC cells. (A) Representative western blot and (B) quantification (normalized to β-actin) of HER2 expression in A549, H1299, H1975, and SK-MES-1 cells 24 h after treatment with 0 or 5 Gy. (C) Relative HER2 mRNA expression in A549 cells determined by real-time quantitative polymerase chain reaction at 0, 0.5, 2, 4, 12, and 24 h following a single fraction of 5 Gy. (D) Representative western blot and (E) quantification (normalized to β-actin) of HER2 expression in A549 cells at 0, 0.5, 1, 2, 4, 12, 24, 48 h, and 7 d post-5 Gy. (F) Representative flow cytometry analysis of cell surface HER2 in A549 cells, with mean fluorescent intensity quantified in (G) for 3 experimental replicates at 0, 0.5, 4, 24, 48 h, and 7 d post-5 Gy. The vertical dotted line in (F) was included to aid in the interpretation of histogram displacement. Data are from 3 independent experiments and presented as mean ± SEM. Statistical significance was determined using unpaired 2-tailed t tests (B) or one-way ANOVA (C, E, G) followed by Dunnett's post hoc test. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Human NSCLC cell lines A549, H1299, H1975, and SK-MES-1 (ATCC: Manassa, VA) were cultured in RPMI-1640 (A549, H1299), ATCC-modified RPMI-1640 (H1975), or DMEM (SK-MES-1) supplemented with 10% fetal bovine serum (FBS) and 1% antibiotic–antimycotic (Wisent Bioproducts: Saint-Jean Baptiste, Canada).

Techniques: Expressing, Western Blot, Real-time Polymerase Chain Reaction, Flow Cytometry