He reduction of en/inv transcript levels, 24 hrs following the 5E1 antibody injections, suggests that the antibody directly, or indirectly, negatively impacted the transcription of this gene. These experiments, however, do not indicate whether lower en/inv transcript levels result from one or both of the en/inv expression domains (the posterior compartment or eyespot centers expression domains; see Fig. 1).Hedgehog’s Role in Wing and Eyespot DevelopmentAdult phenotypes resulting from 5E1 antibody and NS1 medium injectionsInjections of 5E1 or NS1 medium performed on one side of the larval body led to symmetrical changes in both left and right wings suggesting that the antibody, once injected, circulates throughout the hemolymph and is able to target both sides of the animal, and probably most tissues. Wing size. B. anynana and J. get BIBS39 coenia adults injected with the 5E1 antibody as larvae had a smaller forewing Naringin height relative to NS1-injected controls (B. anynana: F1, 82 = 8.62, p = 0.004; J. coenia F1, 140 = 4.47, p = 0.036) (Fig. 5A, B). J. coenia, where we additionally measured wing area, had a smaller forewing area as well (F1, 140 = 6.48, p = 0.012) (Fig. 5C). Wing height reductions in B. anynana were due to the compound effect of reductions across all wing compartments as there were no specific compartments that were more affected than others (Table 1). These compartmentspecific investigations were not undertaken in J. coenia, but the wings appeared also proportionately reduced across the anteriorposterior axis as in B. anynana. Absolute eyespot size. 5E1 injected butterflies had overall smaller eyespots than NS1 injected butterflies, but while some differences were significant others were not. In B. anynana the diameter of the black and gold ring of scales of the largest wing eyespot, the Cu1 forewing eyespot on both dorsal and ventral surfaces, was significantly smaller in 5E1- relative to NS1-injected butterflies (Table 2). In J. coenia most eyespots had some trait that was significantly smaller in 5E1-injected individuals relative to controls. This included all eyespot traits measured on the ventral hindwing and dorsal forewing, as well as the white center of the Cu1 eyespot on the ventral forewing, and both measurements for the Cu1 eyespot on the dorsal hindwing 1317923 (Table 2). Relative eyespot size. The reductions in absolute eyespot size obtained for 5E1-injected butterflies could be due to eyespot differentiation processes having allometrically adjusted to the overall smaller wings. In order to test whether the 5E1 antibody had effects on eyespot size that were independent of its effects on wing size, we performed analyses of co-variance on eyespot traits, corrected for overall wing size (wing height). There was a significant interaction between treatment and wing height for J. coenia’s largest eyespot traits, the diameter of the black and gold ring of the Cu1 forewing eyespot on both dorsal and ventral surfaces, but no such interaction in B. anynana 1379592 (Table 3). The converging (non-parallel) regression lines (Fig. 5G) indicate that smaller wings displayed disproportionately smaller eyespots for the 5E1 treatment relative to the NS1 treatment in J. coenia, whereas treatment had no apparent effect on eyespot size on larger wings in J. coenia (Fig. 5G). This result may simply indicate that stronger effects were seen on smaller animals where the concentration of the antibody was effectively higher (given that the same antibody amount was.He reduction of en/inv transcript levels, 24 hrs following the 5E1 antibody injections, suggests that the antibody directly, or indirectly, negatively impacted the transcription of this gene. These experiments, however, do not indicate whether lower en/inv transcript levels result from one or both of the en/inv expression domains (the posterior compartment or eyespot centers expression domains; see Fig. 1).Hedgehog’s Role in Wing and Eyespot DevelopmentAdult phenotypes resulting from 5E1 antibody and NS1 medium injectionsInjections of 5E1 or NS1 medium performed on one side of the larval body led to symmetrical changes in both left and right wings suggesting that the antibody, once injected, circulates throughout the hemolymph and is able to target both sides of the animal, and probably most tissues. Wing size. B. anynana and J. coenia adults injected with the 5E1 antibody as larvae had a smaller forewing height relative to NS1-injected controls (B. anynana: F1, 82 = 8.62, p = 0.004; J. coenia F1, 140 = 4.47, p = 0.036) (Fig. 5A, B). J. coenia, where we additionally measured wing area, had a smaller forewing area as well (F1, 140 = 6.48, p = 0.012) (Fig. 5C). Wing height reductions in B. anynana were due to the compound effect of reductions across all wing compartments as there were no specific compartments that were more affected than others (Table 1). These compartmentspecific investigations were not undertaken in J. coenia, but the wings appeared also proportionately reduced across the anteriorposterior axis as in B. anynana. Absolute eyespot size. 5E1 injected butterflies had overall smaller eyespots than NS1 injected butterflies, but while some differences were significant others were not. In B. anynana the diameter of the black and gold ring of scales of the largest wing eyespot, the Cu1 forewing eyespot on both dorsal and ventral surfaces, was significantly smaller in 5E1- relative to NS1-injected butterflies (Table 2). In J. coenia most eyespots had some trait that was significantly smaller in 5E1-injected individuals relative to controls. This included all eyespot traits measured on the ventral hindwing and dorsal forewing, as well as the white center of the Cu1 eyespot on the ventral forewing, and both measurements for the Cu1 eyespot on the dorsal hindwing 1317923 (Table 2). Relative eyespot size. The reductions in absolute eyespot size obtained for 5E1-injected butterflies could be due to eyespot differentiation processes having allometrically adjusted to the overall smaller wings. In order to test whether the 5E1 antibody had effects on eyespot size that were independent of its effects on wing size, we performed analyses of co-variance on eyespot traits, corrected for overall wing size (wing height). There was a significant interaction between treatment and wing height for J. coenia’s largest eyespot traits, the diameter of the black and gold ring of the Cu1 forewing eyespot on both dorsal and ventral surfaces, but no such interaction in B. anynana 1379592 (Table 3). The converging (non-parallel) regression lines (Fig. 5G) indicate that smaller wings displayed disproportionately smaller eyespots for the 5E1 treatment relative to the NS1 treatment in J. coenia, whereas treatment had no apparent effect on eyespot size on larger wings in J. coenia (Fig. 5G). This result may simply indicate that stronger effects were seen on smaller animals where the concentration of the antibody was effectively higher (given that the same antibody amount was.
