Fig. S7

RetroMyelin regulates myelination in frog, related to Figure 7 (A) Analysis of overlapping peaks arising from control (left) or RetroMyelin gRNAs-injected Xenopus (right) using CRISP-ID.64 Shown is representative chromatogram from Sanger sequencing of PCR amplicons from genomic DNA extracted from control (left) or RetroMyelin gRNA-injected Xenopus tadpole (right). Alignments of sequences were generated from mixture sequences present in the same animal and compared with the reference sequence (Ref) for the target region. The direction of sgRNA binding site shown in red arrow. CCC (surround by a red rectangle) is the PAM sequence. Three sequences are detected in the injected animal, a: wild-type sequence, b: sequence with nucleotides mutations around the sgRNA target sequence, c: sequence with nucleotides mutations around the sgRNA and 5-bp nucleotides deletions). (B) Longitudinal Z-series of wholemount across the brain stem of Tg(mbp:gfp-ntr) control (upper) and RetroMyelin gRNAs-injected (RNLTR12−/−) (lower) Xenopus tadpoles triply labeled for GFP (green), MBP (red), and XMM (white) (Xenopus myelin marker [XMM]: mAb recognizes specifically an unknown Xenopus myelin protein89). Scale bars, 23 μm. Quantification of oligodendrocytes (GFP+ cells) was plotted. p > 0.05, Mann-Whitney test (two tailed), n = 4 (control), 5 (RetroMyelin gRNA) different sections, mean ± SEM. Intensity of XMM and MBP were quantified and plotted relative to control. ∗p < 0.05, Student’s t test (unpaired and two tailed), n = 4 (control), 5 (RetroMyelin gRNA) different sections.