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Additional Supporting Information may be found in the online version of this article.

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dvdy23964-sup-0001-suppfig1.tif6102KSupplementary figure S1: Analysis of two hearts with b-galactosidase clones in subpopulations of right or left lineage. The heart 7010 presents large clones in the right side of the septum including the RBB (A-B) and in the left ventricle but not in the left Purkinje fibers (LPF). This profile may correspond to a subclone of right lineage. The heart 29001 possesses large clones in the left ventricle including LPF, and in the interventricular septum (IVS) but not in the LBB. This profile may correspond to a subclone of left lineage. Supplementary figure S2: Expression of the Mlc1v-24 and Mlc3f-2E transgenes in the developing ventricular conduction system at fetal stages. Anti-β-galactosidase immunofluorescence on cryosections of foetal hearts at E14.5 (A-B) or E16.5 (C-D) from double transgenic Mlc1v-24:Cx40-GFP (A-C) or Mlc3f-2E:Cx40-GFP hearts (B-D).
dvdy23964-sup-0002-suppfig2.tif4242KSupplementary figure S2 : Expression of the Mlc1v-24 and Mlc3f-2E transgenes in the developing ventricular conduction system at fetal stages. Anti-galactosidase immunofluorescence on cryosections of foetal hearts at E14.5 (A-B) or E16.5 (C-D) from double transgenic Mlc1v-24:Cx40-GFP (A-C) or Mlc3f-2E:Cx40-GFP hearts (B-D).
dvdy23964-sup-0003-suppfig3.tif3861KSupplementary figure S3: Regionalized clusters in the AVN in two smaller clusters. Immunohistochemistry at the level of the AVN using an anti-GFP (A-B) or anti-HCN4 (A'-B') antibody showing β-galactosidase positive cells exclusively in the GFP-negative (A-B) HCN4-positive (A'-B') region of the node.
dvdy23964-sup-0004-suppfig4.tif5297KSupplementary figure S4: Mesp1-Cre genetic tracing analysis in a triple transgenic Mesp1-Cre/Cx40-GFP/R26R embryonic heart. A- Mesp1-derived cells detected by X-gal staining are present in almost all cardiomyocytes including conduction system in brown (Cx40-GFP) and endothelial cells of E14.5 heart. B- The arrow indicates a region of the left ventricular wall negative for X-gal staining and for Cx40-GFP suggesting that Mesp1 non expressing cells participate to the left ventricular myocardium.

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