Fig. 5.

Hedgehog (Hh) pathway activation does not increase TRC differentiation in SOX2 organoids. (A) Hh pathway genes are differentially expressed in SOX2 progenitor populations. (B) Gli1 (green) is expressed by SOX2 IF cells (magenta) outside of taste buds (dashed ovals) (B′,B″, arrows). Image is a confocal z-stack projection. Area outlined in B is shown in B′,B″. (C) Organoids were treated with smoothened agonist (SAG) on days 6-12. See Materials and Methods for details. (D) The hedgehog target gene Gli1 and Sox2 are significantly upregulated by SAG. (E) SAG leads to increased expression of a type I TRC marker gene (Kcnj1) but has little effect on type II (Gnat3) and type III (Pkd2l1) TRC markers. (F) SAG has minimal impact on proliferation (Mki67) in any organoid population. Data are mean±s.e.m. *P<0.05, **P<0.01, ***P<0.001, ****P<0.0001 (one-way ANOVA). (D-F) n=8 (two or three samples each from three independently derived organoid experiments). Scale bar: 10 µm.