Figure 1.

Workflow of MS-based quantitative proteomics during neuronal differentiation. Differentiation of iPSCs toward iN cells was performed using doxycycline-induced expression of Ngn2. Differentiation of MNs was performed using the action of small molecules for neural induction and cell fate determination. Proteins extracted at 10 time points from 2 biological replicates were digested and tandem mass tag (TMT) 10-plex labeled. Peptides were combined and fractionated using high-pH fractionation. The resulting fractions were analyzed by high-resolution nano-LC–MS/MS, and quantification was achieved using TMT 10-plex isobaric labeling.