From mcbyangn@XXXX Thu Feb 27  02:40:21  2003
Envelope-to: djs93@XXXX
Subject: Re: Queries for FlyBase Curation.
Dear David,
Here are my answers in bold:
>Firstly, could you check that the following are correct:
>
>sna/esg/wor triple mutants = sna1; esgG66 double homozygous embryos,
>injected with wor RNAi
san/esg/wor/=sna1, esgG66B double homozygous embryos injected with wor
RNAi.
>sna/esg/wor/pins embryos = sna1,esgG66 double homozygous, that are
>also maternally and zygotically homozygous for rapsP62, injected with
>wor RNAi.
sna/esg/wor/pins embryos = Df(2L)TE35BC-3; pins62
>pins mutant embryos = Embryos both maternally and and zygotically
>homozygous for rapsP62 or rapsP89.
It should be rapsP89 in our paper. But both alleles are null and showed
identical phenotype.
>Secondly, on pg.55 you write:
>
>'In the majority of metaphase pins NBs ectopically expressing Galphai
>(66%, n = 40), DaPKC remains localized as an apical crescent, although the
>intensity is weak compared to wt (not shown). When zygotic Pins is present
>under the same conditions (embryos derived from the same cross), the
>frequency of symmetric NB divisions increases dramatically (82%, n=48).'
>
>Am I correct to infer that the pins mutant neuroblasts were both
>maternally and zygotically homozygous mutant for Pins. The other progeny
>you also clarify which pins (Raps) allele you used for this experiment?
Yes. The allele used in this experiment is pins89.
>Finally, it is not clear to me whether you used sca-Gal4 or neur-Gal4 in
>the following cases:
>
>'When C-terminal Pins (C-Pins) containing the GoLoco motifs (Yu et al.,
>2002 ) are overexpressed in wt embryos...' Pg 55
For this experiment, C-Pins is under the control of a heatshock promoter.
>'In the majority of metaphase pins NBs ectopically expressing Galphai'. Pg 55
For this experiment, Galphai ectopic expression was driven by the sca-gal4.
>'To ascertain the role for Pins in the symmetric NB divisions caused by
>Galphai overexpression, we overexpressed Galphai in pins mutant NBs' Pg 55
Driven by the sca-gal4.
>'To explore this possibility, we expressed Pins-C-Pon in insc mutant
>embryos' Pg 56
Driven by the sca-gal4.
>'When we examined telophase pI ectopically expressing Insc after labeling
>the centrosomes' Pg 57
Driven by the sca-gal4.
>'When Insc is ectopically expressed in ventral epithelial cells of the
>neuroectoderm, Pins/Galphai and Insc/Baz localize to the apical cortex of
>the epithelial cell'
>Pg 58
Driven by the sca-gal4.
Please let me know if you other questions.
Cheers, Xiaohang