DEV035758 Supplementary Material

Files in this Data Supplement:

• Supplemental Figure S1 -

  Fig. S1. PP2A can inhibit larval brain neuroblast overgrowth. (A) mts²⁹⁹/mts^XE-2258 trans-heterozygotes show a neuroblast overproliferation phenotype at 72 hours ALH. (B) Delocalization of aPKC, Numb and Pon in mts²⁹⁹/mts^S5286 mutants is fully rescued by a wild-type mts transgene. (C) PP2A-29B, the A subunit of PP2A, can inhibit excess self-renewal of larval brain neuroblasts. PP2A-29B^RS/PP2A-29B^EP trans-heterozygotes display a neuroblast overproliferation phenotype. (D) tws⁶⁰ larval brains show an overproliferation of neuroblast defects. (E) Quantification of central brain neuroblast numbers in wild type, and in PP2A-29B⁻ and tws⁶⁰ mutants. (F) Increased apoptosis in tws⁶⁰ compared to wild-type larval brains. Apoptotic cells are labeled by anti-activated-Caspase 3 antibody in red. Note that small Dpn-positive cells are observed in tws⁶⁰ mutants, some of which are labeled by activated Caspase. Dotted lines indicate the margins between central brains (to the left) and optic lobes. Scale bars: 10 µm in A for A,C,D,F; 1 µm in B.

• Supplemental Figure S2 -

  Fig. S2. Mts has dual functions as a brain tumor-suppressor and cell cycle regulator. BrdU incorporation was carried out in mts RNAi larval brains expressed under an Insc-Gal4 driver at various temperatures. Gal4 activity is higher at higher temperatures than at lower temperatures. mts RNAi brains have more BrdU labeling at 13°C and 18°C, whereas there is much less BrdU labeling at 25°C or 30°C compared with wild type. Dotted lines indicate the margins between central brains (to the left) and optic lobes. Scale bar: 10 µm.

• Supplemental Figure S3 -

  Fig. S3. Par1 is not required for the regulation of neuroblast self-renewal. par1Δ16 (null allele) MARCM clones (A) and overexpression of Par1 (B) do not show any defects in self-renewal of central brain neuroblasts. Dotted lines label the MARCM clones in A and separate central brains with optic lobes in B. Scale bars: 10 µm.

• Supplemental Figure S4 -

  Fig. S4. aPKC⁰⁶⁴⁰³ significantly suppresses neuroblast overgrowth in mts²⁹⁹. A loss-of-function mutation, aPKC⁰⁶⁴⁰³, partially suppresses the neuroblast overproliferation phenotype of mts larval brains. Double mutant mts²⁹⁹ aPKC⁰⁶⁴⁰³ larvae have many fewer neuroblasts compared with mts²⁹⁹ mutants at 68 hours ALH. Scale bar: 10 µm.