Fig. 2

Knockdown of crbn or cul4 Impairs Head Development

(A) Schematic structure of CRL4^CRBN binding to thalidomide.

(B) Immunoblot analysis (left) and RT-PCR analysis (right) of 24-hpf embryos injected with the indicated MOs against crbn. The indicated antibodies were used to analyze embryos injected with the translation-blocking MO. RT-PCR was performed using the indicated primer set to analyze embryos injected with the splice-blocking MO. The target site of the MO is indicated with red bar in the diagram.

(C and E) One-cell-stage embryos were left uninjected or injected with the indicated MOs against crbn (C) or cul4 (E) with or without corresponding mRNA and then subjected to immunostaining with anti-acetylated α-tubulin antibody at 24 hpf. Fluorescence images (lower panels) and those overlaid with bright-field images (upper panels) are shown.

(D and F) The ratios of head thickness to body length of embryos that were left uninjected or injected with MOs against crbn (D) or cul4 (F) are shown as means ± SD (n = 15 per group). Scale bar, 50 µm. ∗∗∗p < 0.001.