Download 1 Materials and Methods For the MARCM analysis, embryos were

Materials and Methods
For the MARCM analysis, embryos were collected over 2 hours and aged for a further
three hours at 25°C. they were then heat shocked 1hour at 37°C and aged at 25°C to third
instar before analysis. Wildtype clones were generated by the cross yw, elav-gal4,hsFLP; FRTG13, tubulin-gal80 x yw; FRTG13,UAS-mCD8-gfp. ham1 homozygous clones
were generated by the following cross yw; E7-2-36, ham1, FRT40A/CyO x elav-gal4,
UAS-mCD8-gfp, hs-FLP; tubulin-gal80, FRT40A/+.
Fig. S1
Injection of ham dsRNA into embryos carrying the MD marker E7-2-36.
ham dsRNAi leads to a protein null. In embryos carrying the MD marker E7-2-36, ham
dsRNAi leads to the substitution of ES by MD neurons. Thus ham dsRNAi phenocopies
the ham1 phenotype and provides evidence that ham1 is a null allele.
A-D: Buffer (A) and ham dsRNA (C) injected embryos at early stage 17. An abdominal
dorsal cluster is illustrated. ELAV:red, labels all sensory neurons, E7-2-36:green, labels
all MD neurons. A: A buffer injected embryo has a wildtype complement of neurons, the
cluster contains 8 MD, and 5 ES neurons. C: In this ham dsRNA injected embryo there
are 11 MD and one ES neuron. B, D: Diagrammatic representations of A and C
respectively. The ES neurons are represented by red filled circles and MD neurons by
yellow filled diamonds.
1
Table S1
Quantification of HAM expression in ham dsRNA injected embryos.
Injection of ham dsRNA into wild-type embryos leads to the complete loss of HAM
protein expression, and hence causes a HAM null. HAM expression in these embryos
was assayed by immunohistochemical staining with anti-HAM antibody at stages 12-14
when in uninjected or buffer-injected embryos HAM expression is at maximum levels.
HAM+
HAM-
ham dsRNA injected
2
79
Buffer-injected
51
0
2