(A) Representative images of DIV21–24 hippocampal neurons expressing GFP, GFP-Triad3A, Scr shRNA, or Triad3-shRNA treated in the presence of 1 μM TTX for 48 h. Surface AMPA receptors (sGluA1) were visualized by live GluA1 antibody labeling and subsequent fixation under non-permeabilized conditions. Scale bars, 10 μm.
(B–C) Quantitative analysis of surface GluA1 labeling on hippocampal neurons expressing GFPTriad3A (B) or Triad3-shRNA (C). Data represent means ± SEM. *p < 0.05, n = 6–23. **p < 0.005, n = 6–23.
(D) mEPSC recordings from DIV17–21 neurons expressing GFP, GFP-Triad3A or Triad3-shRNA treated with 1 μM TTX for 48 h. Note the increase in mEPSC amplitudes in GFP-expressing neurons upon TTX treatment that is prevented by Triad3A-shRNA and occluded by Triad3A overexpression.
(E) Cumulative mEPSC amplitude distribution summarizing data from GFP (top), GFP-Triad3A (middle), and Triad3-shRNA (bottom) expressing cells in control and following TTX treatment.
(F) Cumulative mEPSC inter-event interval distribution summarizing data from GFP (top), GFPTriad3A (middle), and Triad3-shRNA (bottom) expressing cells in control and following TTX treatment. Note that mEPSC frequency is not affected by GFP-Triad3A or Triad3-shRNA.
(G) Data represent means ± SEM of mEPSC amplitudes. *p < 0.05, n = 11–13.
(H) Data represent means ± SEM of mEPSC frequencies, n = 11–13.