Fig. 2
Blocker sensitivity and distribution of Nav channels. Patch clamp recordings were performed on mature hESC-derived RPE monolayers. a Applying TTX extracellularly (either 1 μM or 10 μM) did not entirely block the current (left). The current was completely removed by intracellular QX-314 (2 mM) (right). Laser scanning confocal microscopy (LSCM) images on Nav distribution in RPE cells. LSCM data inverted greyscale Z-maximum intensity projections of b hESC-derived and c mouse RPE stained against Nav channels (green) and RPE marker CRALBP (red). Scale bars 10 μm. d Immunogold labeling and transmission electron microscopy images showing Nav distribution at the apical membrane in the vicinity of the cell-cell junctions (black arrows). Scale bars 250 nm. e Dissociated hESC-derived RPE cells were let to adhere to poly-l-lysine coated coverslips for 30 min, fixed and immunolabeled against Nav together with CRALBP (up) or tight junction marker ZO-1 (down). The Nav label concentrated on the belt-like region in the middle of the cell, between the basal and apical sides. Scale bars 5 μm