Fig. 5: UNC80’s N-terminal half interacts with NALCN.
a Co-immunoprecipitation assays to locate fragments on UNC80 required for its association with NALCN. Upper panels, schematic presentation of mouse UNC80 truncation mutants used in the studies. Lower panels, HEK293T cells were co-transfected with NALCN and GFP-tagged full-length or truncated UNC80 containing residues as indicated. Cell lysates were immunoprecipitated (IP) with anti-GFP followed by immunoblotting with anti-GFP (lower) or anti-NALCN (top). GFP was used in “ctrl”. It migrated at ~20 kDa (outside the molecular ranges shown) and is not visible in the blots. More than three independent repeats were performed with similar results. b INALCN from cells transfected with NALCN and wild-type or truncated UNC80 mutants as indicated. Recordings were done using a ramp protocol from −100 to +100 mV in 1 s (holding voltage Vh = 0 mV). To ensure that the current was not from nonspecific leak, bath cations were replaced with large non-permeant ion NMDG after each recording (see “Methods” for details). Bar graphs show averaged INALCN amplitude (at −100 mV). Two sample t test of each group against the “full-length” group (n = 11) was performed and the p values are: “Mock” (n = 5, p ≤ 0.001), “1–2387” (n = 9, p ≤ 0.001), “1–2554” (n = 5, p = 0.019), “1–2885” (n = 6, p = 0.818), “1–3000” (n = 5, p = 0.933). Asterisk “*” indicates p < 0.05. Data are presented as mean values ± SEM. Numbers of cells are in parentheses. Source data are provided as a Source Data file.
