Antibodies were affinity-purified on the same epitope as was used for immunization and tested by Western blotting from wild-type and null material or by IP to confirm their specificity (Additional file 5: Figure S3 and Additional file 6: Supplementary methods).
The present results using Akt1-null materials confirmed a previous report by Fujita et al. that gain- and loss-of-functions of PI3K/Akt signaling regulate the DNA binding and transactivity of Runx2 in cultures of osteoblastic cells [27].
Although chic05205 and chic221 are well established strong loss-of-function mutant alleles, only chic221 is a molecularly confirmed null allele (Materials and Methods).
Indeed, wild-type levels of Fz2 in the muscle are not sufficient to rescue the bouton defects induced by presynaptic expression of RNAi-fz2 (Fig. 2A,B,G), yet overexpression of fz2 in the muscle can restore the bouton integrity of fz2 nulls (supplementary material Fig. S1Q) (Mathew et al., 2005).
Moreover, expression of exogenous Delg primarily in the fat body complemented the lethality of delg null alleles (see Materials and Methods).
Their node and edge percolated maps are shown in Fig. 2 (see also Table S1 in Supporting Information) as compared to their maximally random counterparts analyzed as null models (see Materials and Methods).
In addition, two ATPalpha CJ were tested in trans with the null alleles (see " Materials and methods").
However, migration rates in netAB Δ (M+Z) embryos (which are both maternal and zygotic null - see Materials and Methods) were not significantly different from netABΔ embryos at stage 12.5 (p = 0.65) or stage 13 (p = 0.30) (Fig. 4G).
Consequently knock-down of DNT1, by RNAi or the hemizygous introduction of a null allele (Supplementary Material, Fig. S1A), had no effect on longevity and locomotor phenotypes in the adult (Supplementary Material, Fig. S1B E).
Thus, the absorption of peptides and amino acids does not appear to be impaired in the null mutants (Supplementary Material, Fig. S3D).
We found clear expression of MMP1 in the nervous system, but no detectable difference between control, dfmr1 50M heterozygous and dfmr1 50M null brains (supplementary material Fig. S1).
