Fig. 2

Experimental validation of reliable separation of the neuronal ENS from the non-neuronal intestinal cell population. a The bar graph (above) and corresponding data in table (below) show the expression levels in FPKM of five selected genes in the two neuronal replicate [GFP-positive_(GFP + ve) a and _b] and the two non-neuronal replicate [GFP-negative_(GFP-ve) a and _b] cell populations. GFP transcript levels show the level of GFP transcript driven from the phox2b transgene. The expression values of phox2bb measure the endogenous transcript levels of the phox2b gene. Expression levels for the three house-keeping genes actb1, gapdh, and hprt1 are also shown. (b) qPCR validation of expression levels of 7 genes comparing the GFP-positive neuronal populations (Orange bars) versus the GFP-negative non-neuronal (blue bars). chn1, elavl3, chata, phox2bb, and syn2a were more highly expressed in the neuronal population, whereas amy2a and fli1a were more highly expressed in the non-neuronal population. Changes in gene expression were calculated using 2^{− ΔΔCT} method with hprt1 as the house- keeping gene. The mean relative expression of the neuronal population was significantly different from that of the non-neuronal population (p-value < 0.01)