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. 2014 Dec 31;6(7):4936–4952. doi: 10.18632/oncotarget.3210

FIGURE 1. Lung tumor spheres exhibited CSC features.

FIGURE 1

(A) The mRNA levels of stemness-related genes in secondary spheres were detected by RT-qPCR and normalized against mRNA levels of parental cells of NSCLC cell lines. (B) The mRNA levels of stemness-related genes in the first and second passages of spheres from H1299 cells were compared with those in monolayer cells. (C) The protein levels of Oct4, Sox2, and Nanog in monolayers (H1299) and sphere (H1299-S) were detected using flow cytometry. (D) Immunofluorescence analysis was performed to detect the expression levels of Oct4, Sox2, Nanog, and CD133 in monolayers (H1299) and spheres (H1299-S). (E) H1299 cells from monolayers (H1299) and spheres (H1299-S) were analyzed for CD44/CD133 double staining by flow cytometry. Control: isotype control. Data represents the mean ± SD of at least 3 independent experiments, *p < 0.05; **p < 0.01.