Figure 4

KO of Anoctamin 4 leads to an abolishment of Ca²⁺-dependent cation conductance in mouse primary RPE cells. (A) Confocal image of wildtype (upper panel) or Ano4-KO (lower panel) primary RPE cells stained with an antibody against Ano4. Scale bar represents 25 µm. (B) Raw currents through primary RPE cells of wildtype (upper panel) or Ano4-KO (lower panel) before and after the application of ionomycin (1 µM). Application is indicated by the bar. (C) Current density-voltage plot before and after the application of ionomycin in WT (black) and Ano4-KO (white) cells. (D) Fold increase of total current density after the application of ionomycin in WT cells (black bar) or Ano4-KO (white bar) cells). (E) Staining of a murine sagittal section of wildtype (1^st column) or Ano4-KO (3^rd column) with an antibody against Ano4. 2^nd and 4^th column show the respective isotype controls (incubation only with the secondary antibody). Nuclei were stained with DAPI. Upper panel: Scale bar represents 50 µm. GCL: ganglion cell layer; ONL: outer nuclear layer; INL: inner nuclear layer; IS: inner segments, OS: outer segments; RPE: retinal pigment epithelium. Lower panel: Magnification of the upper panel in the RPE. Scale bar represents 10 µm. The values are given as mean ± SEM. ^*p < 0.05.