Fig 5. S-Lap1-HA^int localizes to the elongated mitochondria.

(A) S-Lap1-HA^int protein (green) co-localizes (arrow) with the elongated mitochondria of spermatids, but not with the mitochondria of the elongating cyst (arrowhead) (stained with Mitotracker (red)). Scale bar: 10 μm. (B) Immunoblots of the SDS-soluble supernatant (sn) and the SDS-resistant pellet (p) fraction of the transgenic S-Lap1-HA^int sperm lysates using HA antibody. S-Lap1-HA^int is mainly present in the insoluble fraction of sperm lysate. (C) Immuno-electron micrograph of a cross-section of transgenic S-Lap1-HA^int elongated spermatids (brown dashed line) shows the occurrence of S-Lap1-HA^int protein inside the major mitochondrial derivative (yellow dashed line), represented by gold particles (black dots, arrowheads). Axoneme (red dashed line) and the minor mitochondrial derivative (green dashed line) are also shown. Scale bar: 0.5 μm. (D-E) Cross-section (D) and longitudinal section (E) of immuno-labeled micrographs show S-Lap1-HA^int protein accumulation inside the major mitochondrial derivative (yellow dashed line) in spermatids from homozygous S-Lap1-HA^int; S-Lap1^Δ7 testes, represented by gold particles (black dots, arrowheads). Spermatids (brown dashed line) and axoneme (red dashed line) are labeled. Scale bars: 0.5 μm. (F) S-Lap1-HA^int localizes to the elongated mitochondria in testis of S-Lap1^Δ7, S-Lap2^MI14597, S-Lap3^MB01319, S-Lap4^MB11296, S-Lap5^Δ14, S-Lap6^MI06848 mutants. Scale bar: 200 μm. (G) Immunoblots of total lysate, (H) the SDS-soluble supernatant (sn) and the SDS-resistant pellet (p) fraction of testis expressing the homozygous S-Lap1-HA^int transgene in the classical mutant backgrounds (S-Lap1^Δ7, S-Lap2^MI14597, S-Lap3^MB01319, S-Lap4^MB11296, S-Lap5^Δ14, S-Lap6^MI06848) using HA antibody. One lane represents 1 testis equivalent protein extract.