Fig. 8
Disease-linked Slc7a5 mutations have attenuated effects on Kv1.2. a Kv1.2 channels were co-expressed with Slc7a5[A246V] or Slc7a5[P375L] (1:1) in ltk-mouse fibroblast cells. Conductance-voltage relationships were gathered as described in Fig. 1 (+Slc7a5[A246V] V1/2 = −10 ± 6 mV, k = 11 ± 3 mV, n = 6; +Slc7a5[P375L] V1/2 = −32 ± 8 mV, k = 16 ± 8 mV, n = 10). Dashed lines illustrate conductance-voltage relationships for WT Kv1.2 (black) and Slc7a5 (green) as a reference. b Representative BRET signals were calculated for mEGFP-tagged Slc7a5 mutants co-expressed with Kv1.2-nanoluc, as described in Fig. 6 (representative data from three experiments). c Current density at +60 mV was measured from Kv1.2 co-expressed with various Slc7a5 mutants, before and after a 30 s train of −120 mV hyperpolarizations to −120 mV. d Cell lysates were probed for Slc7a5 expression using western blot for cells transfected with Kv1.2 plus each of the Slc7a5 mutations. No significant changes were detected for expression of the Slc7a5 mutants (n = 3, p > 0.05). e–g Cortical and hippocampal neurons from P2 rat pups transiently transfected at 7 days in vitro with either mCherry-Slc7a5 WT or mCherry-Slc7a5[A246V] or [P375L], and at 10 days in vitro, fixed with 4% paraformaldehyde, permeabilized with 1% Triton X-100 and stained with anti-MAP2 primary antibodies and fluorescent secondary antibodies. Images are representative of three neuronal cultures. Scale bars represent 50 µM
