Survey
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
* Your assessment is very important for improving the workof artificial intelligence, which forms the content of this project
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