Fig. 4

Fig. 4 Expression studies of mab21l2 in wild-type, CE14 and non-CE crispant Xenopus tropicalis.

A Quantitative RT-PCR analysis showing mab21l2 expression levels in wild-type animals through different developmental stages: NF st9 (n = 6), st10.5 (n = 6), st12.5 (n = 6), st18 (n = 5), st20 (n = 5), st23 (n = 6), st28 (n = 6) and st40 (n = 5). Each data point represents mean ± SD. B Histograms demonstrating significantly reduced expression of mab21l2 mRNA in mab21l2-CE14 crispants compared with both non-CE crispants and wild-type embryos at NF stage 20 (CE14 versus non-CE: p = 0.0166, n = 6; CE14 versus wild-type: p = 0.0499, n = 6). In contrast, lrba expression was not affected (CE14 versus non-CE: p = 0.5211, n = 6; CE14 versus wild-type p = 0.3164, n = 6). For both genes, statistical significance was determined using one-way ANOVA followed by Tukey’s post-test. Bars represent mean ± SEM; *p ≤ 0.05. C Whole mount in situ hybridisation throughout different developmental stages in wild-type animals showing mab21l2 mRNA expression in the optic field (blue arrowhead), the lens placode (yellow arrowhead) and a region in the midbrain (red arrowhead). Expression becomes apparent in the hindbrain (H) and the migrating neural crest cells (NC) later in development. Embryos are shown in a lateral view, anterior to the left. D In situ hybridisation showing expression of mab21l2 mRNA in wild-type and crispant embryos at NF stage 18/19. Crispant embryos were injected at the 2-cell stage unilaterally on the right with mab21l2-CE14 or non-CE gRNAs. Frontal, left and right views are depicted, with the left view representing the uninjected side. Patterns of mab21l2 expression in the optic vesicle, lens placode or midbrain (blue, yellow and red arrowheads) are absent or reduced on the injected side for mab21l2-CE14 crispants (upper panel) compared to non-CE crispants (middle panel) and wild-type (lower panel). Source data for panels 4A and 4B are provided in the Source Data file.