Fig. 1: TMEM16K knockout mice.

a RT-PCR from liver and brain tissues obtained from the wild type and TMEM16K full knockout mice. Two different sets of primers amplifying TMEM16K were used, and Gadph and β-actin were amplified as controls. b Representative images and quantification of neuromuscular junction (NMJ) at 6 months of age from wild type (n = 133 NMJ, three animals) and TMEM16K full KO (n = 132 NMJ, four animals) littermates visualized with the fluorescently labeled α-Bungarotoxin. Scale bar 5 µm, Single factor ANOVA, p value = 2.10E−06*** c Representative images and quantification of neuromuscular junction at 24 months of age from WT (n = 137 NMJ, five animals) and TMEM16K full KO (n = 126 NMJ, four animals) littermates. Scale bar 5 µm, Single factor ANOVA, p value = 1.31E−22*** d Representative images of hindlimb clasping, of WT and TMEM16K full KO littermates at 6 and 24 months of age. e Quantification of hindlimb clasping (p value = 0.00003***) and ledge walking (p value = 0.05*) at 24 months of age of neuron specific TMEM16K KO (n = 6 animals) and their WT (n = 7 animals) mice. One-tailed t-test. See Supplementary Movie 1. f Columns 1–3: Immunocytochemistry of U-2OS cells transfected with TMEM16K tagged with V5 epitope and stained for ER-markers Calreticulin, Protein Disulfide-Isomerase (PDI) and Sec61β, respectively. Scale bar 10 µm. g Column 4: Snapshot from live imagining of COS-7 cells expressing with TMEM16K tagged with mClover3 and ER-tdTomato. Scale bar 2 µm. See Supplementary Movie 2. i Quantification of the colocalization of TMEM16K and ER-markers Calreticulin, PDI and Sec61β using Mander’s overlap coefficient, as well as quantification of the colocalization of Sec61β with Calreticulin and PDI measured in the same manner. Sec61β cololicalization with other ER markers is included to provide a meaningful context for the colocalization analysis with TMEM16K, given that Sec61β is a pore forming component of the translocon complex localized exclusively to the ER (three biological replicates, n = 38 cells for TMEM16K vs PDI, 44 for TMEM16K vs Calreticulin, 20 for TMEM16K vs Sec61β, 42 for Sec61β vs Calreticulin, 38 for Sec61β vs PDI). In the box and whiskers plot, the box includes the first quartile and the third quartile, with the central line representing the median. Whiskers represent the minimum and maximum values of data. X inside the box represents the mean of data. Source data are provided as a Source Data file.