Fig. 5
From: The flipflop orphan genes are required for limb bud eversion in the Tribolium embryo
Marker gene expression and apoptosis analysis in Tc-ff RNAi embryos. Ventral focal plane: a, b, b´´, b, d, e-f´. dorsal focal plane: b´, d´. a-b´´: Tc-drumstick; c-d´: Tc-Distalless; e-f´: apoptosis-marker anti-Dcp1. (a, b-b″) Tc-drumstick (Tc-drm; TC006347) marker gene expression in embryonic wildtype (WT) and Tc-ff RNAi embryos marks the segmental base of appendages. (b, b′) Inverted mandible clearly displays wildtype Tc-drm expression at the dorsal focal plane (b′). (c, d, d’) Tc-Distalless (Tc-Dll) marker gene expression in embryonic wildtype (c) and Tc-ff RNAi embryos (d, d’) marks the distal portion of each appendage except for the mandible. (d) Some leg anlagen are not visible in the ventral focal plane (circles) but are visible as inverted limb buds in the dorsal focal plane (arrows) (d’). (e-f) Apoptosis marker anti-Dcp1 in wildtype (e) and Tc-ff RNAi embryos (f-f′). (d) Inverted appendage buds do not show elevated cell death at the stage of bud formation (circles). (f′) Nuclear staining (DAPI) visualises embryonic morphology of inverted appendages after Tc-ff knockdown. A1 abdominal segment 1; Lb labium; Md mandible; Mx maxilla; T1 thoracic segment 1; scale bar 100 μm; all panels in all pictures: anterior to the left