Fig. 3

Downregulation of NHE9 expression via miR-135a affects U87 cell proliferation and migration. a Proliferation of U87 cells transfected with miR-Scr or miR-135a were determined by MTS assay. Absorbance at 490 nm for each sample was normalized to the samples absorbance on day 1. b Loss of proliferative ability in miR-135a transfected U87 glioma cells is reversed by ectopic expression of NHE9-GFP. Proliferation determination and normalization were done as described in (a). c Representative images of U87 cell migration assay taken at 0.5,16 and 26 h. Migration of U87 cells into cell free zones (red dashed-circles) was detected using live cell nuclear counterstain NucBlue (white dots). Loss of migration capacity in miR-135a transfected U87 glioma cells is reversed by ectopic expression of NHE9-GFP. Scale bar (yellow line) is 250 μm. (d and e) Graphs represent normalized fluorescence intensity from NucBlue staining of live cells migrating into the cell free zones at various time points as indicated. Fluorescence intensity of each sample was normalized to the samples fluorescence at 0.5 h. Error bars represent standard deviation (SD); **p < 0.01. Statistical analysis was done using student’s t-test. Graph represents an average of at least three biological replicates