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Fig. 4. Markers for distinct cardioblast sub-types reveal a requirement of tin for cardial cell diversifications. Dorsal views of stage 15-16 wild-type (left column) and tin-ABD, tin³⁴⁶ mutant embryos (right column). (A,B) Staining with antibodies against Mef2 (green) and Doc2+3 proteins (red/red arrowheads). Doc2+3 expands into all cardioblasts in mutant embryos that lack tin expression in the dorsal vessel. (C) Ladybird (Lbe) is detected in two cardioblasts per hemisegment (green arrowheads) just posterior to the Doc-positive pair in the wild type. (D) Cardiac Lbe expression is absent in tin-ABD, tin mutants except for the outflow region (^*). (E) In normal stage 16 embryos, Sur mRNA is detected at high levels in Tin cardioblasts and at very low levels in Doc⁺ cells (red arrowheads). (F) Sur mRNA is barely detectable in mutants lacking cardiac tin expression. (G) In the wild type, ß3-Tubulin (ß3-Tub) is detected in Tin⁺/Doc^- cells of the dorsal vessel (bracket) and in somatic muscles. (H) ß3-Tubulin is maintained with a slightly irregular segmental pattern (bracket) in cardioblasts of tin-ABD, tin³⁴⁶ mutants. (I,J) Detection of svp mRNA and Tin protein. svp is normally expressed in the Tin-negative cardioblasts (I, green arrowheads). In tin-ABD, tin mutant embryos (J) svp expression retains its normal pattern (red arrow indicates sporadic pericardial cell expression of Tin in tin-ABD, tin mutants; rg, ring gland).

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