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. 2016 Dec 23;24(3):469–480. doi: 10.1038/cdd.2016.143

Figure 6.

Figure 6

Slug and Snail are required for Pellino-1-mediated increased migration in A549 cells. Pellino-1 expression is correlated with Slug and Snail expression levels in human lung adenocarcinoma tissues. (a and b) A549 cells were transfected with Myc-Pellino-1 in combination with shLuc, shSlug#1, shSlug#2 (a) or shSnail#1, shSnail#2 (b). At 36 h post-transfection, the cells were harvested and subjected to immunoblotting for Myc, Slug, Snail, E-cadherin, Vimentin and actin. (c and d) A549 cells were transfected with Myc-Pellino-1 in combination with shLuc, shSlug#1, shSlug#2 (c) or shSnail#1, shSnail#2 (d) and subsequently subjected to transwell migration assay. Representative images are shown. The scale bars represent 100 μm. The data are presented as the means±S.D. of at least three independent experiments. The P-values shown were calculated using unpaired Student's t-test. *P<0.05; **P<0.01; ***P<0.005. (e) Representative IHC images for Pellino-1 and Slug expression in lung adenocarcinoma patients who showed strong expression for both Pellino-1 and Slug (top) versus those showing no expression for both proteins (bottom). (f) The patients (n=169) were classified into two groups according to the following Pellino-1 and Slug expression levels: for Pellino-1, those with score 0 versus score 1–3, and for Slug, those with score 0–2 versus score 3. Patients with Pellino-1 expression showed higher Slug expression with statistical significance (P=0.016 by Pearson χ2 test). (g) Representative IHC images for Pellino-1 and Snail expression from lung adenocarcinoma patients who showed strong expression for both Pellino-1 and Snail (left) versus those showing little expression for both proteins (right) are shown. (h) A significantly strong positive correlation between the Pellino-1 and Snail IHC scores was observed (Spearman's Rho=0.445, P=0.000). (i) Patients (n=86) were classified into two groups according to the following Pellino-1 and Snail expression levels: for Pellino-1, those with score 0 versus score 1–3, and for Snail, those with score 0–1 versus score 2–3. Patients with Pellino-1 expression showed significantly higher Slug expression (P=0.000 by Pearson χ2 test)