Supplementary Figure Legends
Figure S1. (a) List of mutational site in the KLF4 gene domain in lung small cell carcinoma
from TCGA database. (b) Luciferase reporter assays testing the effect of KLF4 mutation on
function of KLF4 in repressing cyclinB1 promoter activity. Data are represented as mean ± SD.
(c) KLF4 expression in normal lung and lung small cell carcinoma from TCGA database. The P
value was calculated based on a linear mixed model.
Figure S2. KLF1-17 expression in normal lung and lung tumor tissues from TCGA
adenocarcinoma samples. P values were calculated based on linear mixed models.
Figure S3. (a) Cell proliferation assay to test the effect of KLF4 in lung epithelial cell line
BEAS-2B. Control: vector-carrying lentivirus infected cells. Data are represented as mean ± SD.
(b) Western blot showing overexpression of KLF4 in KLF4-carrying-lentivirus-infected cells
(KLF4) compared with vector-lentivirus-infected cells (control).
Figure S4. Expression of cell markers in the mouse lung. (a-b) IHC staining for Cav1 in normal
(a) and Klf4-deleted (b) mouse lung. (c-d) IHC staining for CC10 in normal (c) and Klf4-deleted
(d) mouse lung. (e-f) IHC staining for FoxJ1 in normal (e) and Klf4-deleted (f) mouse lung. Scale
bars for (a-b) and (e-f): 30 m. Scale bars for (c-d): 50 m.
Figure S5. (a) Correlation between KLF4 expression and methylation in lung cancer patients
from TCGA lung small cell carcinoma database. (b) mRNA expression of HDACs in normal
lung and lung tumor tissues from TCGA adenocarcinoma samples. P values were calculated
based on linear mixed models.
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Figure S6. (a) Luciferase reporter assay in 293T cells testing the effect of HDAC1-6 on KLF4
promoter activities. “*”, P < 0.05. (b) Luciferase reporter assays testing the effect of HDAC3 on
promoter activity of KLF4 in A549lung cancer cell line. Data are represented as mean ± SD.
Figure S7. H&E analysis of the mouse lung 16 weeks post infection. (a) H&E staining of lung
tissue from wildtype mice (contrl), K-rasLSL-G12D/+ mice, Klf4fl/fl mice, and K-rasLSL-G12D/+;Klf4fl/fl
mice by 16 weeks post infection. Scale bars from left to right: 2 mm, 200 m, and 100 m. (b)
Statistical analysis of average numbers of nodules in each lung lobe from the four types of mice
by 16 weeks post infection.
Figure S8. IHC staining of mouse lung tissue for Bmi1. Scale bars from left to right: 2 mm, 200
m, and 100 m.
Figure S9. IHC staining of mouse lung tissue for Myeloperoxidase. Scale bars from left to right:
2 mm and 100 m.
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