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The reasons for reduced courtship rates of the ML line males cannot be resolved here, but our present results and competitive fitness [1] data nonetheless suggest that tester females found them more attractive than control males, accepting similar numbers of mating from ML males despite their reduced duration of courtship.
The origin of the femur is chosen as the mid-point of the ML line.
From the midpoint of the ML line, two additional lines were drawn at 60° in the posteromedial (PM line) and posterolateral directions (PL line).
This line was then shifted in the anterior-posterior direction until the widest medial to lateral portion of the tibia was located, which was termed the ML line.
The tubes were then filled to the 5 mL line with sterile water, capped, sealed with Parafilm, and kept at room temperature in the dark for one day.
In order to generate enough microsporidian material to start the evolution experiment, material from an infected beetle population was split evenly between 5 standard 50 ml tubes (Sarstedt, Nümbrecht, Germany), the 50 ml tubes filled up to the 15 ml line with a mixture of flour (Alnatura type 550) plus 5 % brewer's yeast and the tubes vortexed thoroughly at multiple angles.
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In our experiments, self-sealing reaction vessels with a volume of approximately 250 mL, lined with gold, and which permit withdrawal of samples at the temperatures and pressures of interest, were used.
The derivation of the male-limited (ML) lines and their matching controls (C) is described in detail in [1].
Figure 3C shows that SMTB 1 44 applied to the column elutes at 1.38 ml (blue line), while PAI-1 elutes at 1.11 ml (green line).
fl, forelimb; hl, hindlimb; ml, milk line; mr, mandibular and maxillary region; mg, mammary gland; hfp, hair follicle placodes.
Clinical samples comprising blood (40 mL), central lines (tips), urine (20 mL), and sputum were collected at 06:00 [ 11].
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