Extended Data Fig. 4: Generation of Tmem63 knock-out flies.
From: Drosophila TMEM63 and mouse TMEM63A are lysosomal mechanosensory ion channels
a, Tmem631 null mutant was generated through CRISPR-Cas9 mediated genome editing. The coding sequence of Tmem63 was replaced with coding sequences of GFP and mini white. The GFP was not expressed in this line possibly due to a deletion in 5’ UTR (shown as dashed lines). Arrowheads indicate cleavage sites of Cas9. b, PCR validation of Tmem631 allele. c, Tmem632 null mutant was generated by ends-out homologous recombination. The coding sequence of Tmem63 was replaced with Gal4 and mini white. The GAL4 was expressed in this line under the control of the native promoter of Tmem63. d, PCR validation of Tmem632 allele. The images in b and d are representatives of three independent experiments. The locations of the primers in b and d are shown in a and c, respectively. The sizes of the PCR fragments are indicated below the DNA bands. Unprocessed gels are available as source data.
