Fig 2: Kaplan–Meier analysis in patients with stage 1 non-small cell lung cancer according to aldehyde dehydrogenase 1A1 (ALDH1A1) and CD133 expression. (A) Recurrence-free survival in ALDH1A1 low versus high expression, (B) overall survival in ALDH1A1 low versus high expression, (C) recurrence-free survival in CD133 low versus high and (D) overall survival in CD133 low versus high expression.

Fig 3: Morphological alterations of GSC-like cells were observed on different days, and expression of tumor stem cell-associated genes was detected by western blotting and RT-qPCR. (A) Glioma cells. (B) GSC-2 days. (C) GSC-3 days. (D) GSC-7 days. (E-G) GSC-P1, P5 and P10. Scale bars, 50 µm. (H) The expression level of CD133 (130 kDa), c-Myc (57-70 kDa), Klf4 (65 kDa), SOX2 (35 kDa), Nanog (42 kDa) and β-actin (42 kDa) was examined by western blotting. (I) Grayscale analysis results of western blotting. (J) CD133, c-Myc, Klf4, SOX2 and Nanog expression at the mRNA level was analyzed by RT-qPCR and normalized to β-actin. *P<0.05 and **P<0.01 vs. U87 as control group. GSC, glioma stem cells; Klf4, Krueppel-like factor 4; P1, passage 1; P5, passage 5; P10, passage 10; d, day; RT-qPCR, reverse transcription-quantitative PCR.

Fig 4: Detection of the effects of M5-5d intervention on GSC characteristics via western blotting and reverse transcription-quantitative PCR. (A and B) The protein expression levels of stem cell-associated genes, CD133 (130 kDa), c-Myc (57-70 kDa), Klf4 (65 kDa), SOX2 (35 kDa), Nestin (207 kDa), Nanog (42 kDa) and OCT4 (45 kDa), were analyzed via western blotting. GAPDH (37 kDa) was used as an internal control. (C) Total RNA was extracted to quantify CD133, c-Myc, Klf4, SOX2, Nestin, Nanog and OCT4 at the mRNA level, which was normalized to GAPDH. *P<0.05, **P<0.01 and ***P<0.001 vs. untreated GSCs as control group. Klf4, Krueppel-like factor 4; M, Mdivi-1; GSC, glioma stem cells.

Fig 5: Representative immunohistochemical staining intensity of CD133 (left column) and aldehyde dehydrogenase 1A1 (ALDH1A1) (right column) from patients with stage 1 non-small cell lung cancer. In all cases, 0=no staining, 1+=mild, 2+=moderate and 3+=strong intensity of staining. Photographs were taken at magnification 200×.