Extended Data Fig. 9: Lysosomal recordings and CRISPR-mediated gene disruption in N2a cells.
From: Drosophila TMEM63 and mouse TMEM63A are lysosomal mechanosensory ion channels
a, Enlarged lysosomes (LAMP1-YFP-positive intracellular vesicles) in N2a cells. Scale bar, 5 µm. b, Schematic of the experimental procedure for patch-clamp recordings on lysosomes of N2a cells. Lysosomes are shown as grey vesicles. N, cell nucleus. c, DNA sequences of the Cas9-targeted segments in wild-type (WT) and MsTmem63a knock-out (KO) N2a cells. Guide RNA and protospacer adjacent motif (PAM) sequences are highlighted in red and blue, respectively. d, The deletion in the MsTmem63a gene results in a pre-mature stop codon of the MsTMEM63A proteins in KO cells. e, The mRNA level of MsTmem63a in WT and KO cells. Data shown are mean ± s.e.m. Statistical significance was determined by two-sided Mann-Whitney test. n = 6 independent experiments. f, The MsTMEM63A protein level in WT and KO cells. The results in a and f are representatives of three independent experiments. Numerical data and unprocessed blots are available as source data.
