Journal: Development (Cambridge, England)
Article Title: Lineage-specific effects of Notch/Numb signaling in post-embryonic development of the Drosophila brain
doi: 10.1242/dev.041699
Figure Lengend Snippet: The adPN Nb makes one PN at one time. (A) The principle of twin-spot MARCM. Following mitotic recombination, one homozygous daughter cell loses repressor 1 (miRNA against rCD2) and becomes specifically labeled with reporter 1 (UAS-rCD2::RFP), while the other daughter cell loses repressor 2 (miRNA against GFP) and expresses reporter 2 (UAS-mCD8::GFP). As to clones derived from a self-renewing Nb, one can label the GMC clone and the remaining lineage in different colors at the same time using twin-spot MARCM. (B-G) Merged confocal images of adult adPN twin-spot MARCM clones generated at early (B,E), middle (C,F), or late (D,G) larval stages. Nb clones of different sizes (green) were consistently accompanied by lone adPNs (magenta). Brain neuropils were counterstained with nc82 mAb (blue). Scale bar: 20 μm.
Article Snippet: Primary antibodies used in this study include rat anti-mCD8 mAb (1:100; Caltag), rabbit anti-GFP Ab (1:1000; Molecular Probes), mouse anti-Acj6 mAb (1:100; DSHB), mouse anti-nc82 mAb (1:100; DSHB), mouse anti-Elav (1:200; DSHB), mouse anti-rCD2 mAb (1:100; Serotec), rat anti-Dpn mAb (1:2) ( Boone and Doe, 2008 ), rabbit anti-pH3 Ab (1:200; Upstate), rabbit anti-Dsred (1:500; Clontech) and rabbit anti-cleaved caspase-3 (1:200; Cell Signaling).
Techniques: Labeling, Clone Assay, Derivative Assay, Generated