Fig. 3

Both WT and mutant LIN28B promote cell invasion and migration in BE2C cells. (A) Western blotting of LIN28B_WT/MU using the BE2C-TET cells untreated or treated with 50 ng/mL doxycycline for 3 d. α-Tubulin was used as a loading control. (B) Relative cell growth of BE2C-TET cells untreated or treated with 50-ng/mL doxycycline. Values represent means ± SEM of triplicate experiments. (C) Schematic of transwell migration and invasion assay. (D and E) Transwell migration and invasion assay of the BE2C-TET cells untreated or treated with 50-ng/mL doxycycline. Cells were stained with 0.1% crystal violet (D), and the number of migrated cells through the membrane per field was compared with the two-tailed unpaired t test (E). Values represent means ± SD of triplicate experiments. ****P < 0.0001. (Scale bar, 100 µm.) (F) qRT-PCR to detect let-7d and let-7i expressions in BE2C-TET cells that were either untreated or treated with 50-ng/mL doxycycline for 3 d. Values were normalized to U6, U47, and RNU44 small nuclear RNAs and represent the means ± SD of triplicate experiments. Statistical analysis was performed using the two-tailed unpaired t test. ****P < 0.0001.