Fig. 3

The negative impact of the biallelic variations in POLR3B. (A) The expression of POLR3B mutants by quantitative real-time PCR. Three separate experiments were performed (nonparametric test; *p < 0.05; error bars, s.e.m.). (B) The Western blot analysis of POLR3B expression in cells transfected with plasmids carrying variants. Three independent experiments were performed. (C) PyMol visualization was performed to compare the native and missense mutant amino acid. WT and mutant residues are in light green. (D) Larvae of WT maintained a normal posture. But the polr3b morphants exhibit greater bending angles and body curvature (chi-square test, ****p < 0.0001). (E) Statistics of the embryos with posture-related morphological defects. (F–J) Disruption of polr3b impairs the development of precursor cerebellar Purkinje cells. Typical images of in situ hybridization labeling Purkinje precursors cells with anti-sense probes against ptf1a. (K–O) Disruption of polr3b reduces cerebellar granule cells. Typical images of in situ hybridization labeling cerebellar granule cells with anti-sense probes against reelin [Colour figure can be viewed at wileyonlinelibrary.com]