NO donor promotes human lung CSC-like phenotypes.
A, H460 cells were treated with NO donor (DPTA NONOate) for 24 h and analyzed for cell viability by MTT assay. Apoptotic and necrotic cell death after the treatment was analyzed by Hoechst 33342/PI co-staining assays. B, percentages of apoptotic and necrotic nuclei in NO-treated cells were analyzed and calculated as relative to the control cells. C, ×10 immunofluorescence images of the treated and nontreated cells stained with Hoechst 33342/PI. After being treated with DPTA NONOate (0–40 μm) for 5 days, H460 (D), H23 (G), and H292 (J) cells were suspended and subjected to spheroid formation assay. Spheroid number H460 (E), H23 (H), H292 (K), and size H460 (F), H23 (I), H292 (L) were analyzed and calculated as relative to the control after 10, 20, and 40 days. Plots are means ± S.D. (n = 3). *, p < 0.05 versus nontreated cells.