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1.
Figure 4

Figure 4. Microarray profiling of NSCLC cell line H1299 stably transfected with SIX3 and empty vector control.. From: Down-Regulation of SIX3 is Associated with Clinical Outcome in Lung Adenocarcinoma.

(a) Partial heatmap plot of hierarchical clustering of genes that were down-regulated by over-expression of SIX3, including S100P and TGFB3 (highlighted by red cycles) is shown. (b) Validation of down-regulation of some representative genes by over-expression of SIX3 using quantitative RT-PCR.

Min-Li Mo, et al. PLoS One. 2013;8(8):e71816.
2.
Figure 6

Figure 6. Kaplan-Meier analyses of overall survival and progression-free survival (PFS) based on the SIX3 expression levels.. From: Down-Regulation of SIX3 is Associated with Clinical Outcome in Lung Adenocarcinoma.

(a) Overall survival in 145 patients with lung adenocarcinomas, (b) Overall survival in 99 Stage I patients, (c) Overall survival in 64 adenocarcinoma patients with BAC features, (d) Overall survival in 81 adenocarcinoma patients without BAC features, (e) Overall survival in 61 Stage II-IV patients, (f) PFS in 145 patients with lung adenocarcinomas.

Min-Li Mo, et al. PLoS One. 2013;8(8):e71816.
3.
Figure 5

Figure 5. shRNA knock-down of SIX3 in NSCLC cells expressing endogenous SIX3.. From: Down-Regulation of SIX3 is Associated with Clinical Outcome in Lung Adenocarcinoma.

(a) Quantitative RT-PCR of SIX3 expression in cell line H1703 stably transfected with control shRNA or shRNAs against SIX3 (shRNA-1 or -2). (b) MTS assay of H1703 cells stably transfected with SIX3 shRNAs. (c) Wound healing assay of H1703 cells stably transfected with SIX3 shRNAs. Pictures were taken under a light microscope (20×).

Min-Li Mo, et al. PLoS One. 2013;8(8):e71816.
4.
Figure 3

Figure 3. SIX3 suppressed proliferation and migration of NSCLC cells.. From: Down-Regulation of SIX3 is Associated with Clinical Outcome in Lung Adenocarcinoma.

(a) Quantitative RT-PCR of SIX3 expression in cell lines H1299 and H322 stably transfected with control or SIX3 expression vector. (b) MTS assay of SIX3 stably transfected cells. (c) Colony formation assay of SIX3 stably transfected cells. (d) Wound healing assay of SIX3 stably transfected H1299 cells. Pictures were taken under a light microscope (20×).

Min-Li Mo, et al. PLoS One. 2013;8(8):e71816.
5.
Figure 1

Figure 1. Methylation in the promoter region caused down-regulation of SIX3 expression in NSCLC cell lines.. From: Down-Regulation of SIX3 is Associated with Clinical Outcome in Lung Adenocarcinoma.

(a) Quantitative RT- PCR of SIX3 expression in NSCLC cell lines and total RNA of normal adult lung tissues was included as a control. (b) Quantitative MSP of the SIX3 promoter region in NSCLC cell lines. Genomic DNA of normal adult lung tissues was included as a control. (c) Semi-quantitative RT-PCR of SIX3 expression after 5 µM 5-aza-2′-deoxycytidine (DAC) treatment in NSCLC cell lines. GAPDH was used as control for RNA quality and loading.

Min-Li Mo, et al. PLoS One. 2013;8(8):e71816.
6.
Figure 2

Figure 2. Methylation in the promoter region caused down-regulation of SIX3 expression in NSCLC tissue samples.. From: Down-Regulation of SIX3 is Associated with Clinical Outcome in Lung Adenocarcinoma.

(a) Quantitative RT-PCR of SIX3 expression in matched pairs of fresh NSCLC tumor tissues and adjacent normal lung tissues, (b) Quantitative MSP of the SIX3 promoter region in matched pairs of fresh NSCLC tumor tissues and adjacent normal lung tissues. (c–f): Immunohistochemistry (IHC) examples of SIX3 protein in matched normal lung tissues (c and e) and tumor tissues (d and f) from two patients (#10 (c and d) and #12 (e and f)). SIX3 protein is represented by positive nuclear staining. Pictures were taken under using a light microscope (magnification 40×).

Min-Li Mo, et al. PLoS One. 2013;8(8):e71816.

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