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. 2021 Aug 23;11:17029. doi: 10.1038/s41598-021-96227-5

Figure 2.

Figure 2

Expression of cortical glutamatergic excitatory markers and tau in iNgn2 neurons. (A) Immunostaining of SMI312 (axonal marker) and MAP2 (dendritic marker) in 10-day iNgn2 neurons. Scale bar represents 50 μm. (B) As assessed by western blots, there is robust expression of synaptic proteins Synapsin 1 and PSD-95, and the glutamatergic marker GluR1 in 14-day iNgn2 neurons from acutely transduced iPSC and NPC, and the iNgn2-NPC stable cell line. For comparison, human brain tissue from the anterior cingulate cortex (ACC) was also probed. Full-length blots are presented in Supplementary Fig. S11. (C) By 18 days of iNgn2, the neurons exhibit numerous synaptic punctae consisting of colocalized presynaptic marker VGLUT1 and postsynaptic marker HOMER1 along neuronal processes immunostained with the p-Tau marker PHF-1. Scale bar represents 20 μm. (D) These neurons also express cortical layer marker and tau mRNA (as transcripts per million, TPM) as determined by RNAseq.