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. 2017 Jun 1;20(6):874–890.e7. doi: 10.1016/j.stem.2017.02.014

Figure 5.

Figure 5

Monitoring the Dynamics of Primed-State to Naive-State PSC Conversion Using Cell Surface Protein Markers

(A) Phase contrast images of H9 PSCs reveal the morphological changes that occur during primed-state to naive-state conversion under t2i/L+PKCi conditions. Doxycycline-inducible NANOG and KLF2 transgenes were activated for the first 8 days in t2i/L, and then doxycycline was withdrawn and PKCi was added. Scale bars, 100 μm.

(B) Flow cytometry dotplots of pairwise antibody combinations over the time course. Shown are primed-specific markers on the y axis (CD57, top; CD24, bottom) and naive-specific markers on the x axis (CD75, top; CD130, bottom).

(C and D) FlowSOM visualization of the flow cytometry time course data for (C) H9 PSCs under t2i/L+PKCi conditions and (D) WIBR3 under 5i/L/A conditions. Note that 5i/L/A conversion is transgene-free and that 5i/L/A was added on day 1. The minimal spanning trees of the self-organizing maps display an unsupervised clustering of the samples based on their cell surface protein expression levels (right). The heatmap shows the expression level of each cell surface protein marker in the cell clusters (left).