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Fig. 3 | BMC Cell Biology

Fig. 3

From: Testis-specific ATP synthase peripheral stalk subunits required for tissue-specific mitochondrial morphogenesis in Drosophila

Fig. 3

Knon-GFP rescues the mutant phenotype and localizes to mitochondria in primary spermatocytes and post-meiotic spermatids. Phase-contrast and paired fluorescence images of cells from testes of knon ms(2)1400 /Df(2R)7E; knon-GFP/+ males, stained with Hoechst. Knon-GFP (green) was not detectable in spermatogonia (a) but was associated with phase-dark mitochondria (red arrows) in all later stages (b-i). Yellow arrows indicate nuclei; red arrows indicate mitochondria-associated Knon-GFP. In primary spermatocytes (b), Knon-GFP-labeled mitochondria were small and diffuse in the cytoplasm. In meiotic cells (c), Knon-GFP-marked mitochondria associated with the spindle; immediately after meiosis (d) mitochondria aggregated beside each nucleus and were associated with a strong Knon-GFP signal. The mitochondrial derivatives within the Nebenkern at the onion stage (e, red arrow) and during early- (f) and mid-elongation (g) showed unambiguous Knon-GFP localization. In nearly mature elongating spermatid cysts (h), elongated flagella appear wild type with no clumped mitochondrial derivatives (compare to Fig. 1). Knon-GFP is evenly distributed throughout the elongated spermatids. After sperm individualization, motile sperm (i) retain detectable Knon-GFP (red arrow). The apparent syncytial nature of some cells at earlier stages is an artefact of the preparation—ring canals connecting cells in a cyst are commonly burst open by the pressure from a cover slip. These localization results are identical to those seen when Knon-GFP is expressed in a wild-type background (not shown). Scale bars in a-g, h, and i are 20 μm

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