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(B – B′, D – F′ ) Btd RNAi knockdown type II NB lineages (B – B′ ) or type II NB clones homozygous mutant for btd XG81 (D – E′ ) or btd XA (F – F′ ) in 3rd instar larval brains produce Ase− immature INPs (open arrows) and a few Ase+ daughter cells (arrows) but no mature INPs.
Although pntp1 mutants by themselves do not affect type II neuroblast identity, double mutant for btd and pntp1 might have more severe phenotype than btd or pntp1 mutants alone.
Type II neuroblasts mutant for btd do not turn on Ase, but their INPs never mature.
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(B) Since btd-GAL4 is inserted in the promoter of btd, we could not do the rescue by using btd-GAL4 to drive the expression of UAS-mSp8 or UAS-btd in type II NB clones homozygous mutant for other btd mutant alleles, such as btd XG81.
For btd loss-of-function analyses, yw btd XA FRT19A/FM7c and yw btd XG81 FRT19A/FM7c (Wimmer et al., 1993; Estella and Mann, 2010) were used for generating btd mutant clones and UAS-btd RNAi (#29453; Bloomington Drosophila stock Center, Bloomington, Indiana) for Btd RNAi knockdown.
UAS-mCD8-GFP driven by Btd-GAL4 was utilized as reporter for btd expression.
Finally, they show that over-expressing btd in type II neuroblasts that are mutant for trx rescues type II neuroblast identity.
Indeed, Pigott et al. [ 33] reported that replacing loops 1 and 2, or 2 and 3 simultaneously with CDR3 peptide mutant toxins maintained the binding affinity for Bt-R1 [ 33].
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