Journal: iScience
Article Title: A translational study on the survival and molecular mechanism of PD-L1 expression in EGFR-mutant NSCLC treated with osimertinib
doi: 10.1016/j.isci.2025.114175
Figure Lengend Snippet: IFN-γ induces PD-L1 expression in EGFR -mutant NSCLC cell lines via STAT3 activation (A) Quantitative real-time PCR analysis of IFNG and PD-L1 gene expression in various lung cancer cell lines. (B–D) Western blot analysis demonstrating PD-L1 expression across cell lines (B), STAT3 phosphorylation following treatment with STAT3 inhibitor C188-9 in PC-9 (left) and HCC827 (right) cells (C), and PD-L1 expression and STAT3 phosphorylation after IFN-γ treatment for 30 min or 18 h in PC-9 (left) and HCC827 (right) cells. Ctrl denotes vehicle control. (E) Flow cytometry analysis of cell surface PD-L1 expression following IFN-γ stimulation (∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001). (F and G) Western blot analysis of PD-L1 expression in PC-9 (F) and HCC827 (G) cells after STAT3 knockdown, STAT3 overexpression, or C188-9 treatment. (H) Flow cytometry analysis of cell surface PD-L1 expression following C188-9 treatment (∗∗∗ p < 0.001). (I and J) Immunofluorescence analysis of STAT3 localization in HCC827 (I) and PC-9 (J) cells that overexpress STAT3 (STAT3-OE) or treated with IFN-γ (IFNG). NC indicates negative control. Nuclei were stained with DAPI. Red boxes in the merge images highlight areas shown at higher magnification in the fourth column. Scale bars: 10 and 2 μm for full and zoom images (data are presented as mean ± SD).
Article Snippet: APC anti-human PD-L1 antibody was purchased from Proteintech.
Techniques: Expressing, Mutagenesis, Activation Assay, Real-time Polymerase Chain Reaction, Gene Expression, Western Blot, Phospho-proteomics, Control, Flow Cytometry, Knockdown, Over Expression, Immunofluorescence, Negative Control, Staining