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Fig. 2 | BMC Biology

Fig. 2

From: Female factors modulate Sex Peptide’s association with sperm in Drosophila melanogaster

Fig. 2

The levels of LTR-SFPs bound to sperm increase from male ejaculate to sperm stored in female seminal receptacle, but the pattern differs from SP’s. Pre-mating ejaculate samples were collected from Fru>dTRPA1 males exposed to high temperatures, as described in “Methods”. Post-mating sperm samples were isolated from wild type (CS) females that had mated to wildtype (CS) males and frozen at 35 min and 2 h ASM. Sperm heads were stained with DAPI (blue) and LTR-SFPs were visualized with Alexa fluor 488, staining the sperm (green); Bar = 20μm. Sperm isolated from male ejaculate immediately after exudation (A, D, G, and J) were probed for CG1656 (degree of freedom, F(2, 42)=19.48), Antares (F(2,30)=20.77), CG1652 (F(2,36)=72.79), and CG9997 (F(2,21)=87.66), respectively. Sperm isolated from mated female bursa, frozen at 35 min ASM (B, E, H, and K) were probed for CG1656, Antares, CG1652, and CG9997, respectively. Sperm isolated from mated female’s seminal receptacle, frozen at 2 h ASM (C, F, I, and L) were probed for CG1656, Antares, CG1652, and CG9997, respectively. The insets show the negative controls for their respective panels. Sperm samples in negative controls were incubated with only secondary antibody (anti-rabbit, Alexa fluor 488) but no primary antibody (anti-LTR-SFP) incubation, as mentioned previously. C’, F’, I’, and L’ show corrected fluorescence (CF) intensity of CG1656, Antares, CG1652, and CG9997 on sperm at three different stages and time points. Error bars show Mean±SE (AU stands for arbitrary units); p***<0.001, ns=not significant, n=8–15

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