Figure 1

Menin inhibits cell migration and represses PTN/RPTP β/ζ expression. (a) Men1^+/+ and Men1^−/− MEFs were added to the upper-filter of a transwell, and 24 h later, the migrated cells were stained with crystal violet, and cell number was counted under a microscope. (b) A549 cells were transfected with either vector-expressing shRNAs against Luc or one of the three shRNAs against MEN1 and selected by G418. The efficiency of menin silencing was determined by western blotting. Equal sample loading was confirmed by Ponceau S staining of the Western blot membrane. (c) The selected A549 cells were added to the upper-filter, and cell migration was determined. (d) Increased RPTP β/ζ mRNA levels in Men1^−/− MEFs were detected by real-time quantitative reverse transcriptase (qRT)–PCR. The amount of RPTP β/ζ mRNA was determined by normalizing their mRNA quantity with the control β-actin mRNA level, and mean values and s.d. were calculated from triplicates of a representative experiment. (e) The efficiency of menin ablation and the effect of Men1 expression on PTN (18 kDa) and RPTP β/ζ (≈250 kDa) expression were determined by western blotting, and β-actin was used as a loading control. Equal sample loading was also confirmed by Ponceau S staining of the western blot membrane. Results in all cases are mean±s.d. percentage change in number of migrating cells vs corresponding untreated cells (default=100) and calculated from triplicates of an independent experiment, *P<0.05 vs control.