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. Author manuscript; available in PMC: 2018 Nov 1.
Published in final edited form as: Alzheimers Dement. 2017 Apr 18;13(11):1237–1250. doi: 10.1016/j.jalz.2017.03.001

Figure 2. Cellular characterization of iPSC-derived cells by quantitative RT-PCR and immunofluorescence.

Figure 2

(A) iPSCs were differentiated into dopaminergic (mDA) and cholinergic (BFCN) neurons and each stage and neuronal type was confirmed using quantitative real-time RT-PCR for cellular specific markers (Supplementary Table S1). Levels of mRNAs were measured by real-time RT-PCR and calculated relatively to the geometric mean of GAPDH-mRNA and PPIA-mRNA reference controls using the 2−ΔΔCT method. Each column represents the mean of two biological and technical replicates. The error bars represent the Standard Error of the Mean (S.E.M.). (B–F) Immunofluorescence (IF) analyses of the cell types across differentiation. (B) The pluripotency markers POU5F1 (red) and NANOG (green) in iPSCs. (C) The NPCs marker, Nestin (red), and the marker expressed by floor plate progenitors, FOXA2 (red) in MD progenitors. (D) The dopaminergic marker, TH (green) and the mature neurons marker, TUBB3 (red) in mDA. Co-localization of the markers indicates the differentiation of mature dopaminergic neurons. (E) The NPCs marker, Nestin (red), and the marker indicating the pattern of basal forebrain cholinergic differentiation, Nkx2.1 (green) in MGE progenitors. (F) The cholinergic marker, ChAT (green) and the mature neuron marker TUBB3 (red) in BFCN. Co-localization of the markers indicates the differentiation of mature BFCN. DNA counterstaining in blue (B–F).