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. 2024 Feb 20;12:1316048. doi: 10.3389/fcell.2024.1316048

FIGURE 5.

FIGURE 5

Prdm15 is required for head development in X. laevis. (A) Unilateral KD of Prdm15 leads to a microcephaly phenotype compared to control MO (CoMO)–injected embryos. Co-injection of human full-length PRDM15-WT RNA rescues the head phenotype. The dorsal views of representative stage 43 embryos are shown. Dashed lines indicate the measured area and length. (B) Statistical evaluation of smaller and deformed heads as indicated in (A) reveals a significantly smaller head area (orange), head width (purple), and interocular distance (light blue), while co-injection of hPRDM15 RNA rescues almost all Prdm15 MO–induced phenotypes. (C) Ventral view of Alcian blue–stained and dissected cranial cartilages from control and Prdm15 morphants (stage 45). Prdm15 KD results in a mild and severe phenotype showing narrowed or deformed cartilage structures (black arrowheads), especially at the branchial arch (ba). (D) Anterior view of CoMO-injected and Prdm15 MO–injected embryos (stage 20) after whole-mount in situ hybridization (WMISH) with NCC-specific marker genes during NCC migration such as snai2, egr2, and twist1 show a reduced marker gene expression on Prdm15 MO–injected side (black arrowheads). (E) Statistical evaluation of NCC-specific marker genes' expression as illustrated in (D) reveals a significant reduction in marker gene expression upon Prdm15 MO KD. (F) Lateral view of CoMO-injected and Prdm15 MO–injected embryos (stage 23) after WMISH with the NCC marker foxd3 shows a reduced expression on the Prdm15 MO–injected side (black arrowhead). Statistical evaluation of foxd3 expression reveals a significant reduction in its expression upon Prdm15 MO KD. Abbreviations: CoMO, control morpholino oligonucleotide; egr2, early growth response 2; foxd3, forkhead box D3; inj., injected; hPRDM15, human PRDM15; MO, morpholino oligonucleotide; n, number of independent experiments; N, number of analyzed embryos in total; ns, non-significant; Prdm15, PR-domain zinc finger protein 15; snai2, snail family transcriptional repressor 2; st., stage; twist1, twist family bHLH transcription factor 1; uninj., un-injected; WMISH, whole-mount in situ hybridization; WT, wild type. Error bars indicate standard errors of the means. ns, p > 0.05; *, p ≤ 0.05; ****, p ≤ 0.0001.