Fig. S3

The developmental and hematopoietic defects observed in the Taf3-depleted embryos are not due to non-specific p53 activation. A: qRT-PCR analysis monitoring levels of p53, 113 p53 (an alternatively spliced isoform of p53 that is known to be highly upregulated in zebrafish following MO off-target effects) (Robu et al., 2007), p21 (a well-characterized p53 target gene), and gapdh in taf3 MO-treated embryos. As a control, expression of p53, 113 p53, and p21 were also monitored in control MO-treated embryos, which was arbitrarily set to 1. Error bars represent SD. The results show that the levels of p53, 113 p53 and p21 mRNAs are not significantly affected in Taf3-depleted embryos, indicating that the hematopoietic defect observed in the Taf3 morphants is not due to non-specific activation of the p53 pathway. B: Whole-mount in situ hybridization with a riboprobe to hbbe1 or gata1 (28 hpf) in embryos injected with a control MO or a taf3 MO, or co-injected with taf3 and p53 MOs. The results show that hbbe1 and gata1 expression in taf3 MO-injected embryos was equivalent to that obtained in embryos co-injected with taf3 and p53 MOs, indicating the hematopoietic defect was not due to p53 activation. C: Acridine orange staining of embryos (30 hpf) injected with a control MO or a taf3 MO, or co-injected with taf3 and p53 MOs. The results show that there was no difference in staining between Taf3 MO-injected embryos and those co-injected with MOs directed against both Taf3 and p53.