Supplementary Figure 8: Correlative image analysis and the mining of KEGG database identified the molecular signaling pathways in the morphogenetic domains (the most anterior position). We identified 20 distinct gene expression patterns by graphics image clustering analysis (clusters shown in the order of left-to-right, top-to-bottom). (A) In the most anterior position, along the A-P axis, we detected the widely expression pattern of Notch signaling pathway for example in cluster 2 (pharynx), cluster 5 (hypodermis and neuron) and cluster 10 (muscle), which indicates that Notching signaling plays the central role in the regulation of cell fate specification and the coordination of multiple cell types in early organogenesis. During the digestive tube morphogenesis, specifically, Notch signaling is required to prevent cross-fusion between pm8 (pharyngeal muscle) and vpi1, the pharyngeal-intestinal valve cell that contributes to the formation of the tube structure linking pharynx to intestine. (B) In the most anterior position, along the V-D axis, the Calcium signaling pathway was detected in cluster 6 (muscle) and cluster 19 (pharynx) but not cluster 9, 14 and 17 (anterior region neuron). The pathway plays the roles in regulating muscle contraction and pharynx pumping, and contributes to control the processes of intercellular communication, germ layer formation and the establishment of body axes during gastrulation. (C) In the most anterior position, along the L-R axis, MAPK signaling pathway is found in cluster 1, 7, 8 and 9 (muscle) but not cluster 2 and 19 (anterior region neuron). mpk-1, the C. elegans homolog of the conserved mitogen activated protein (MAP) kinase ERK, is the repressor of par-1 gene that is required for the control of cell polarity and asymmetric division. Loss of function of mpk-1 restores the viability of par-1 mutant with corrected phenotypes in asynchronous cell divisions, and the asymmetric distribution of cell fate specification markers [35].