Figure 5. PM2.5 elevates the proliferation in NSCLC cells. (A–E) Th17 cells were treated with 100 μg/cm2 of PM2.5 for 24 h, and then all cells and supernatants were collected for the analysis. (A) TNF-α and IL-6 levels in the supernatants were measured using ELISA. (B) IL-17a contents in the collected supernatants were calculated using ELISA. (C) RT-qPCR and (D) western blot analysis were used to measure IL-17a expression levels in the harvested cells. (E) IF staining of IL-17a in the harvested cells. Scale bar, 20 μm. (F) Th17 cells were treated with 100 μg/cm2 of PM2.5 for 24 h, and then the conditional medium was collected, and mixed with fresh RPMI1640 absolute medium at 1:3. The composed culture medium was exposed to A549 and H1350 cells for 12, 24, 48 or 72 h. Then, the NSCLC cells were collected for cell proliferation analysis using CCK-8 analysis. (G, H) Th17 cells were incubated with 100 μg/cm2 of PM2.5 for 24 h, and then the conditional medium was collected, and mixed with fresh RPMI1640 absolute medium at 1:3. Then, the composed culture medium was subjected to A549 and H1350 cells for another 24 h. Subsequently, all cells were harvested to assess the cell proliferation using colony formation and EdU assays. Scale bar, 100 μm. Quantification of colony formation assay and EdU was exhibited. All data are expressed as mean ± SEM. *p<0.05 and **p<0.01 compared to the Ctrl group.