Fig. S3

Knockdown of bcdin3 and larp7 rescues nucleotide stress-induced phenotypes in the neural crest where hexim1 is expressed, related to Figure 3.

(A) Western blot for hexim1 and β-actin (actb) in uninjected and 8 ng of hexim1 MO injected embryos at 24 hpf.

(B) Two 600 bp in situ hybridization probes to the mature hexim1 transcript were designed.

(C) Both antisense probes and their corresponding sense controls were tested on 18-somite stage zebrafish embryos (~20 embryos per condition).

(D) Zebrafish hexim1 expression at various developmental stages with probe 1 (~20 embryos per stage). Results are identical if probe 2 is used.

(E) Zebrafish embryos were injected with 10 ng of bcdin3 or 12 ng of larp7 MOs that were previously published. Embryos were treated with either DMSO or 6.5 µM leflunomide at 50% epiboly and subjected to in situ hybridization at 24 hpf for crestin and mitfa expression. Dorsal views of embryos are shown. Numbers indicate the number of embryos with the shown phenotype vs. the total number of embryos in the clutch.

(F) Embryos treated as described in (E) were also scored for melanocytes at 2 dpf.

(G) Real time RT-PCR validation of HEXIM1, HEXIM2 and GAPDH knockdown in the A375 cells by pooled siRNAs (mean of 3 replicates ± SD). Data is normalized to ACTB.