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(A) Schematic showing horizontal view of E12 olfactory epithelium (OlfE). (B,C) similar sections of E12 OlfE with control pro-neural gene Ascl1 and PpG gene Sox2 . Ascl1 is expressed in scattered cells, often with colocalized 3’UTR/CDS expression, while Sox2 shows graded differential 3’UTR/CDS expression across OlfE regions. * denotes symmetrical patterns in contralateral OlfE for each gene, (additional genes in S2B, S2D-S2H Fig in S1 File ). (D-G) higher power view of core PpG expression in OlfE SVZ; as labeled. Note that all PpGs (but not Ascl1 (D)) show preferential localization of their 3’UTR sequences to the apical plate, labeled in D and same in E-G. Basal plate shows high Nestin (D,E) and Sox2 (F,G) protein and each CDS can also be colocalized to Nestin positive radial glia (seen at longer exposure) (green carrots). Apical stem cell zone is marked. For Myc both the 3’UTR and CDS are concentrated apically. For all other PPGs examined, Sox2 , Oct4 , Klf4 , and Nanog , their 3’UTR expression is concentrated apically with the CDS signal both apical and extending along radial glia to the basal plate; carrots; F,G and S2 Fig in S1 File ). (H) Schematic of whisker key anatomy. (I,J) low power views of four developing whiskers; top two with well-defined dermal papillae, bottom two- earlier condensates. Note that differential 3’UTR/CDS expression for all PpGs (I, J and S2M, S2N Fig in S1 File ) is apparent and an early indicator of hair peg condensation. (K-M) higher power views showing differential 3’UTR/CDS for Sox2 (L) and Myc (M) in anatomically distinct regions of the whisker. Shh (K) shows coincidental 3’UTR/CDS expression in an adjacent section (carrot). Other PgPs shown in S2I-S2L Fig in S1 File ). Anatomical abbreviations are, Shh; Shh expression region, DP; dermal papilla, DS; dermal sheath (white). Scale bar, A; 500, B,C; 350, D-G; 150, H,K-M; 55, I,J; 350 um. |