A 3D reconstruction of the cardiac outflow tract (OFT) region (horizontal and vertical rotation) from stage-16 wild-type embryo stained for Ladybird early (Lbe; blue) and Tinman (Tin; red). Heart-anchoring cells (HANCs) are not shown. Note that at this stage in the tip of aorta, three pairs of Tin-expressing cardioblasts lie anteriorly to Lbe/Tin-positive cardioblasts (light pink). The most anterior Tin-expressing cardioblasts correspond to the pair of anterior cardioblasts (ACBs) initially associated with cardiac outflow muscles (COMs) (refer to Fig. 1). Movie is SI Movie 3 in Proc Natl Acad Sci USA (2008) 105: 2475-2480. Movies from this publication include CIL# 25536, 25537, 25538, 25539, 25540, 25541.
Stage 16 embryos were stained with primary antibodies: mouse anti-Lbe, rabbit anti-Tin. Species-specific secondary antibodies were conjugated to Alexa Fluor 488, Cy3 or Cy5. The double stained embryos were visualized using a Zeiss LSM 510 Meta confocal microscope. Images representing 0.5 mm optical sections were compiled and processed by Volocity software (Improvision).
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