WNT inhibition elevates olfactory placode expression and increases the number of neural progenitors. (A) qRT-PCR results at day 24 for olfactory placode (POU2F1, DLX5, EMX2) and neural progenitor (ASCL1, SOX2). Values are normalized to 18S and compared with vehicle-treated group (n = 5, mean ± SEM). (B) Immunocytochemistry analysis comparing protein expression on day 24 of differentiation for POU2F1 (green) and DLX5 (red). Scale bars, 100 μM. (C) Quantification of POU2F1- and DLX5-positive cells. Values are normalized to DAPI and compared with vehicle-treated group (n = 5, mean ± SEM). (D) Immunocytochemistry analysis comparing protein expression on day 24 of differentiation for ASCL1 (green) and SOX2 (red). Scale bars, 100 μM. (E) Quantification of ASCL1- and SOX2-positive cells. Values are normalized to DAPI and compared with vehicle-treated group (n = 5, mean ± SEM). P-values (*P < 0.05, **P < 0.01, ***P < 0.001). WNT, wingless/integrated proteins.