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A–B’’
Immunostaining of wild‐type (A–A’’) or nos‐Gal/UASp‐CCR4‐HA (B–B’’) germaria with anti‐Aub (red), and anti‐CCR4 or anti‐HA (green). GSCs are shown in (A–A’’). White arrowheads in (B–B’’) indicate cytoplasmic accumulation of CCR4‐HA colocalized with Aub in GSCs.
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C
Co‐immunoprecipitation (IP) of NOT1, NOT3, and CCR4 with GFP‐Aub in ovaries. Wild‐type (WT, mock IP) or nos‐Gal4/UASp‐GFP‐Aub (GFP‐Aub) ovarian extracts were immunoprecipitated with anti‐GFP, either in the absence or the presence of RNase A. Western blots were revealed with anti‐GFP, anti‐NOT1, anti‐NOT3, and anti‐CCR4. Inputs are extracts prior to IP.
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D
Co‐IP of NOT1 and NOT3 with GFP‐Aub or GFP‐AubAA in ovaries. Wild‐type (WT, mock IP), nos‐Gal4/UASp‐GFP‐Aub (GFP‐Aub), or nos‐Gal4/UASp‐GFP‐Aub
AA (GFP‐AubAA) ovarian extracts were immunoprecipitated with anti‐GFP in the presence of RNase A. Western blots were revealed with anti‐GFP, anti‐NOT1, and anti‐NOT3. Inputs are extracts prior to IP.
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E–G
Genetic interaction between aub and twin in GSC self‐renewal. Immunostaining of germaria with anti‐Vasa (green) and anti‐Hts (red). DAPI (blue) was used to visualize DNA. Examples of aub
HN2/+, twin
DG24102 and aub
HN2/+; twin
DG24102 germaria are shown. White arrowheads indicate GSCs; the white arrow indicates GSC loss.
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H
Quantification of mutant germaria with no GSC in 3‐, 7‐, and 14‐day‐old females of the genotypes shown in (D–F). The number of scored germaria (n) is indicated. ***P‐value < 0.001 using the χ2 test.
Data information: Scale bars: 10 μm in (A–B’’) and (E–G).
