Fig. 4.

miR135a acidifies sorting endosomes by downregulating NHE9. a NHE9-GFP colocalizes with Rab5 in sorting endosomes of U87 cells as determined by immunofluorescence microscopy. Top panel: NHE9-GFP (green) Middle panel: sorting endosome marker, Rab5 (red) and Bottom panel: Merge. Colocalization is indicated by yellow in the merge. Scale bar is 10 μm. Quantification of NHE9-GFP localization with Rab5 in U87 cells was done using Manders’ coefficient (0.51 ± 0.05. n = 50 cells). b Calibration of endosomal pH in U87 cells (c) pH in sorting endosomes is acidified in U87 cells transfected with miR-135a relative to scrambled control. Graph represents mean from three biological replicates and at least 50 cells were used for pH quantification in each experiment. Error bars represent standard deviation (SD); *p < 0.05. Statistical analysis was done using student’s t-test