Supplementary Figure 1. lgl and pins regulate larval neuroblast numbers.
a-d, 96h ALH larval brains stained for the indicated markers. a, wild type. b,c, lgl334 and dlgm52
mutants have an increase in Miranda+ neuroblasts and Prospero+ GMCs. d, pins62 mutants have
the opposite phenotype.
Supplementary Figure 2. Apoptosis is not increased in pins mutant brains.
a,b, 96h ALH larval brains stained for the apoptosis marker activated caspase-3 (Casp-3) and the
neuroblast marker Miranda. Both wild type (a) and pins62 mutant (b) brains have similar levels of
apoptosis.
Supplementary Figure 3. A comparison of neuroblast and GMC markers reveals that lgl
pins mutant brains have supernumerary neuroblasts.
a, Schematic of wild type neuroblast progeny showing that neuroblasts transcribe worniu,
deadpan, miranda, and prospero (Broadus et al., 1998, Nature 391:792; Figure 2; data not
shown); maintain proliferation (take up BrdU; Figure 2); and fail to differentiate (never express
the neuronal differentiation marker Elav; Figure 2). In contrast, GMCs do not transcribe any of
the above-mentioned genes (Broadus et al., 1998, Nature 391:792; Figure 2; data not shown); exit
the cell cycle (fail to take up BrdU); and intitiate differentiation into Elav+ neurons. prospero is
transcribed in neuroblasts, translated in neuroblasts, and partitioned into GMCs (Broadus et al.,
1998, Nature 391:792).
b, Schematic of lgl pins neuroblast progeny showing that both daughter cells transcribe worniu,
deadpan, miranda, and prospero; maintain proliferation (take up BrdU); and fail to differentiate
(never express the neuronal differentiation marker Elav) (Broadus et al., 1998, Nature 391:792;
Figure 2; data not shown). prospero is transcribed and translated in both daughter cells but is
unable to be partitioned out of one daughter cell due to lack of cortical polarity machinery (Figure
3f). Thus, the proliferating cells in lgl pins brains are neuroblasts by molecular, cellular, and
developmental criteria.
Supplementary Figure 4. aPKC is required for neuroblast self-renewal in lgl mutants.
Heterozygosity for aPKCk06403 reduced ectopic neuroblast formation in lgl334 mutants at all stages
examined (a) and in multiple lgl mutant genotypes (b). *, wild type pupariates by 120h ALH.