Figure 3

Figure 3

CHTF18 patient variants fail to support T cell development.(A) Effect of chtf18 knockdown on T cell development at 5 days post fertilization (dpf) in zebrafish embryos. T cell development was assessed by whole-mount in situ hybridization (WISH) using a probe for the T cell specific kinase, lck. Numbers on images represent the fractions of embryos with the depicted phenotype. Thymus is indicated by dashed blue ovals. (right panel) The efficacy of chtf18 E2I2 splice-blocking MO (1ng) was assessed by RT-PCR at 1 dpf. (B) Capacity of patient CHTF18 variants to rescue T cell development in zebrafish. Lateral images of zebrafish treated with control or chtf18 MO and subjected to rescue by heat-induced re-expression of the WT or mutant human CHTF18 orthologs. Rescue of T-cell development was evaluated by lck WISH at 5 dpf. Thymus staining is indicated by dashed blue ovals and the fraction of embryos with the depicted phenotype is indicated on each micrograph. The intensity of the lck WISH signal was quantified by ImageJ software and is depicted graphically as box and whisker plots. Statistical significance was evaluated using one way ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001.