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Fig. 3 | BMC Biology

Fig. 3

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

Fig. 3

Ablated SSCs in Send1>Rh1G69D females affects neither the binding of SP to sperm (at 2 h ASM) nor the long-term gradual cleavage of SP from sperm (at 4 days or 8 days ASM). Expression of ER stress-inducing Rh1G69D by spermathecae-specific Send1-Gal4 results in ablation of SSCs. A,B Control (Send1>CyO) females mated with ProtB-eGFP males (eGFP-tagged sperm; green) show normal numbers of SSCs (stained with DAPI; white dotted circle) lining the spermathecal cap (red dotted circle). C,D Experimental (Send1>Rh1G69D) females mated with ProtB-eGFP males show ablated SSCs (stained with DAPI) lining the spermathecal cap (red dotted circle); n=10; Bar = 20μm. E Western blot probed for SP and indicated LTR- SFPs at 2 h ASM. Lanes# 1: Fv, reproductive tract (RT) of 4 unmated females (negative control), 2: MAG, 1 pair of male accessory glands (positive control), 3: C, sperm dissected from SR of 30 control (Send1>CyO) females mated to wild type (CS) males at 2 h ASM, 4: Exp, sperm dissected from SR of 30 experimental (Send1>Rh1G69D) females mated to wild type (CS) males at 2 h ASM. Lanes were probed for SP and LTR-SFPs CG1656, CG1652, Antares, and CG9997 as described in the text. F Western blot probed for SP at 4 and 8 days ASM. Lanes# 1: Fv, reproductive tract (RT) of 4 unmated females (negative control), 2: MAG, 1 pair of male accessory glands (positive control), 3: C, sperm dissected from SR of 30 control (Send1>CyO) females mated to wild type (CS) males at 4 days ASM, 4: Exp, sperm dissected from SR of 30 experimental (Send1>Rh1G69D) females mated to wild type (CS) males at 4 days ASM, 5: C, sperm dissected from SR of 30 control (Send1>CyO) females mated to wild type (CS) males at 8 days ASM, 6: Exp, sperm dissected from SR of 30 experimental (Send1>Rh1G69D) females mated to wild type (CS) males at 8 days ASM. Lanes were probed for SP. Tubulin served as loading control

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