Sanger sequencing of the Mp3g19030.1 gene in Mp<i>thar</i> mutants and amino acid alignment of RTN4IP1 homologues.

(A, B, and C) Regions of the Mp3g19030.1 gene in Mpthar2 (A), Mpthar4 (B), and Mpthar6 (C) were Sanger sequenced to confirm the SNPs identified by next generation sequencing. The consensus sequence is shown on the top rows, and results from the Sanger sequencing are shown on the second rows. The blue arrows indicate the site of mutation in each Mpthar mutant, and the tick marks above the sequences indicate the position of the nucleotide in the Mp3g19030.1 gene. (D) Trimmed amino acid alignment of RTN4IP1 homologues. Protein sequences similar to HsRTN4IP1 from 10 species were identified using the BlastP algorithm to search their proteomes [37]. The sequences were aligned via the L-INS-i strategy in MAFFT [76] and manually trimmed. (TIF)