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. 2018 Feb 5;217(2):571–583. doi: 10.1083/jcb.201706101

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© 2018 Kavaler et al.

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Figure 5. — miR-279/996 facilitates the activity of nuclear Notch in the PNS lineage. (A–E) Images of adult nota. (A) Wild-type fly shows the normal pattern of external mechanosensory bristles. In each heminotum, there are five rows of small microchaete bristles from the center of the notum to the dorsocentral macrochaete (large) bristles. (B) The mir-279/996 null mutant exhibits mostly normal exterior bristle patterning, but some macrochaetes are lost (asterisks). Notch heterozygote also exhibits mostly normal bristle patterning (C), whereas double heterozygote of Notch and mir-279/996 exhibits a mild increase in microchaete density (D); arrow points at an extra partial row of bristles. (E) Notch heterozygote in the mir-279/996 null shows extreme loss of exterior bristle structures. (F–K) Shown are pupal nota at >28 h APF stained for cell-specific markers. (F and G) Normal pattern of DPax2 (large shaft and small sheath nuclei), Pros (sheath nuclei), and Elav (neuronal nuclei) in control w[1118] (F) and N[55E11]/+ (G). (H) N[55E11]+; mir-279/996[15C/15C] notum shows loss of most DPax2 and Pros staining and development of “all-Elav” sensory organ lineages. Thus, the exterior balding of the adult flies of this genotype (E) is caused by conversion of most lineage cells into neurons. (I–K) Pupal nota that were subjected to a 2 h heat shock during pIIIb division. (I) An appropriately timed pulse of hs-NICD results in relatively uniform transformation of neurons into sheath cells. (J) Heat shock treatment of mir-279/996 null animals shows the typical sheath-to-neuron phenotype of this mutant. (K) Parallel treatment of hs-NICD, mir-279/996 null animals reveals a large population of double neuron organs (dotted circles) and/or lineages that coexpress Pros with detectable Elav reactivity (arrowheads). A rare case of double Pros/double Elav organ is indicated with double lines. Epistasis of the miRNA phenotype indicates that NICD cannot function effectively without miR-279/996. Bars, 50 µM.