Fig. 6.

Loss-of-function of either CREB or Rab11b impairs TGF-β2-mediated KCC2 trafficking to the plasma membrane. Immunofluorescence for KCC2 in primary mouse hippocampal neurons transiently transfected with control negative siRNA (neg siRNA, green in B and F), with specific siRNA against CREB1 (si CREB, green in C and G) or with specific siRNA against Rab11b (si Rab11b, green in D and H). Cells shown in A and E represent non-transfected neurons. At 24 h following transfection cells were treated with 2 ng/ml TGF-β2 for 60 min. Nuclei were labeled with DAPI (blue). Scale bar: 10 µm. Asterisks indicate cytosolic KCC2 localization, white arrows point to plasma membrane labeling. The KCC2 distribution profile was visualized by line scans for KCC2 immunofluorescence. Black arrows indicate peaks of KCC2 immunoreactivity. Representative images and line scans are from four independent experiments.