Figure 2
MiR-133α/AFTPH axis regulated NTR1 trafficking to plasma membrane during recovery in human colonic epithelial cells.
(A) NCM460-NTR1 cells transfected with as-miR-133α and as-miR-control were exposed to NT (100 nM, 1 hr), washed and replenished with NT-free media (recovery). Trafficking of NTR1 to plasma membrane was attenuated in miR-133α knocked-down NCM460-NTR1 cells, as visualized using immunocytochemistry. (arrow: membrane-bound; arrowhead: vesicle-bound) Scale bars, 10 μm (B) Reduced membrane-bound NTR1 was present in the miR-133α-silenced cells after recovery when compared with control cells, as measured in biotinylation assay. *p < 0.05 when compared to as-miR-control-transfected cells. (C) NCM460-NTR1 (NTR1) cells and cells overexpressing NTR1 and AFTPH (NTR1-AFTPH) were exposed to NT, washed and allowed to recover as described in (A). Trafficking of NTR1 to plasma membrane was attenuated in NCM460-NTR1-AFTPH cells as visualized using immunocytochemistry when compared with NCM460-NTR1 cells. (arrow: membrane-bound; arrowhead: vesicle-bound) Scale bars, 10 μm (D) Reduced membrane-bound NTR1 was observed after recovery in NCM460-NTR1-AFTPH cells when compared to NCM460-NTR1 cells in biotinylation assay. *p < 0.05 when compared to NCM460-NTR1 cells.
