Drosophila Ataxin 2-binding protein 1 marks an intermediate step in the molecular differentiation of female germline cysts

DEV050575 Supplementary Material

Files in this Data Supplement:

• Supplemental Table S1 -
• Supplemental Figure S1 -

  Fig. S1. The GFP Protein trap in A2BP1^CC00511 reflects the expression and localization of the endogenous protein. (A) A germarium from an A2BP1^CC00511 female stained with anti-A2BP1 (red), anti-GFP (green) and 1B1 (blue). The pattern and localization of GFP and A2BP1 are virtually identical. (B) In situ hybridization showing A2BP1 RNA expression.

• Supplemental Figure S2 -

  Fig. S2. Disruption of A2BP1 function leads to extra mitotic divisions resulting in egg chambers with 31 nurse cells. (A,A′) w¹¹¹⁸ and (B,B′) A2BP1^CC00511 hemizygous egg chambers stained with DAPI and phalloidin. (C) w¹¹¹⁸ and (D) A2BP1^CC00511/Df(3L)ED4457 ovaries stained with Bam (green) and alpha-Spectrin (red). The Bam expression domain is expanded in the A2BP1^CC00511/Df(3L)ED4457 background. (E) Graph showing quantification of the phenotype and suppression by a bam-null mutation. (F) Introduction of an encore^q4 mutation into the A2BP1^CC00511/Df(3L)ED4457 background results in the persistence of fusome material in egg chambers. A2BP1^CC00511/Df(3L)ED4457 ovaries and (G) encore^q4/+; A2BP1^CC00511/Df(3L)ED4457 ovaries stained with 1B1 (red) and DAPI (blue). (H) Graph showing quantification of the phenotype and enhancement by the encore^q4 mutation.

• Supplemental Figure S3 -

  Fig. S3. Disruption of A2BP1 does not result in overlapping expression between Nanos and Bam. Control and A2BP1^e03440 hemizygous germaria stained for Nanos (green) and Bam (red). The early expression of both Nanos and Bam do not appear to be disrupted in A2BP1^e03440 mutants. The Bam expression domain appears to be greatly expanded in the A2BP1 mutant background.

• Supplemental Figure S4 -

  Fig. S4. Different A2BP1 isoforms interact with one another. (A,B) Ovarioles stained for Vasa (green), 1B1 (red) and DAPI (blue). (A) A2BP1^KG06463/A2BP1^f01889. (B) A2BP1^KG06463/ A2BP1^e03440. The A2BP1^KG06463allele in combination with A2BP1^f01889or A2BP1^e03440exhibits germ cell counting defects, fusome perdurance past when the organelle is normally degraded and an occasional tumorous phenotype. (C) Western blot showing A2BP1 levels in ovaries from w¹¹¹⁸, A2BP1^CC00511 homozygotes and various mutant backgrounds, including other A2BP1 alleles and rbp9²⁶⁹⁰. rbp9²⁶⁹⁰ mutants display a tumorous phenotype but express normal levels of A2BP1. The membrane is blotted with polyclonal guinea pig anti-A2BP1 antibody at 1:5000. Red arrows indicate A2BP1 isoforms that exhibit different expression levels in different mutants and green arrows indicate the shifted A2BP1 isoforms due to GFP fusion in the A2BP1^CC00511 line. (D) Yeast two-hybrid assay showing the interaction between two different A2BP1 isoforms, RE and RI. (E) Co-immunoprecipitation from S2 cells showing that A2BP1 isoforms bind to themselves.