Skip to main content
. 2020 Aug 18;161(10):bqaa141. doi: 10.1210/endocr/bqaa141

Figure 2.

Figure 2.

Definitive endoderm differentiation was improved with the inclusion of the cell cycle inhibitor PD0332991. Cells were stained with SSEA-4 at day 0 of differentiation (A, top plot) and at day 5 (A, bottom plot). Histograms show SSEA-4 expressing cells (red) with gates determined based on unstained cells (blue). (B) The CDK4/6 inhibitor PD0332991 was added to medium as described in Fig. 1 or was absent during DE differentiation. Those cells were then analyzed for RNA expression of FOXA2 and SOX17 at the end of DE differentiation (n = 4 independent experiments). Some cells subjected to ±PD0332991 were reserved for cytospins and immunofluorescently stained with anti-SOX17 and DAPI. Green (SOX17) and blue (DAPI) cells from + or - PD0332991 were counted and graphed in C, with P < 0.0001. Images for SOX17 staining are shown in D with -PD0332991 on the top and +PD0332991 on the bottom. White scalebar represents 25 μm. Error bars on graphs are standard error of the mean (SEM). DE, definitive endoderm; SSEA4, stage-specific embryonic antigen-4.