Figure 3
Expression patterns of BgLpR in the fat body and the ovary of B. germanica. The study was carried out in adult females during the 7 days of the first gonadotrophic cycle. 7c means 7-day-old adult females with basal oocytes chorionated. A) BgLpR mRNA expression in fat body and ovary analyzed by RT-PCR/Southern blotting. B. germanica actin5C levels were used as a reference. B) The antibody raised against BgLpR recognized a single band in fat body extracts subjected to 7.5% SDS-PAGE under denaturing conditions. C) RNAi experiments to further assess the antibody specificity for BgLpR. Left panel: BgLpR mRNA levels in the fat body of dsControl and dsBgLpR-treated specimens; actin5C levels were used as a reference. Right panel: Western blot analysis using the antibody against BgLpR in the fat body of dsControl and dsBgLpR-treated specimens; the antibody reveals a single band having between 97 and 116 kDa, as in B, corresponding to BgLpR protein. D) Western-blot analysis showing the BgLpR protein pattern in the fat body and the ovary. In the case of fat body, 10 μg of total proteins were loaded in each lane. In that of ovaries, 0.02 ovary equivalents were loaded in each lane. In the later gel, we loaded low amounts of tissue and we exposed the film longer than usual to avoid the "pressing" effect of the huge vitellin band placed just above that of LpR. In films that are exposed even longer, LpR can be also detectable on the days 0, 1 and 2. Blots shown are representative of three replicates.