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. 2015 Dec 15;7(42):67986–68001. doi: 10.18632/oncotarget.6622

Figure 5. SJ26 repressed WNT1 expression and reduced the migration activity of H1299 cells.

Figure 5

A. SJ26 reduced the WNT1 mRNA level. The H1299 cells was incubated with 1, 3, 10, or 30 μM SJ26 for 24 h and then subjected to quantitative RT-PCR analysis. The GAPDH expression level was used as an internal control. The mRNA level of WNT1 expression in untreated H1299 cells was defined as 100%. Results were obtained from the average of three independent experiments. Asterisks indicate p < 0.05. B. SJ26 reduced the Wnt1 protein and its down strean mediator β-catenin levels. H1299 cells were incubated with 1, 3, 10, or 30 μM SJ26 for 24 h. Total cell extracts were prepared, and immunoblotting analysis was conducted using antibodies against Wnt1, β-catenin, or α-tubulin. C. SJ26 reduced the healing activity of H1299 cells. H1299 cells were incubated with 10 μM DMSO or 1, 3, or 10 μM SJ26 for 48 h, and then the treated cells were subjected for scratch assays. Relative migration rates of H1299 cells were determined by migration distance over the time. Cell growth was also determined using the MTT assay. The value of migration rate or cell growth in DMSO-treated cells was defined as 100%. Asterisks indicate p < 0.05. D. WNT1-overexpression reversed the healing-inhibitory activity of SJ26. The relative healing rates were determined in 10 μM SJ26-treated cells that overexpressing WNT1. The relative healing rate in DMSO-treated H1299 cells was defined as 100%.