Skip to main content
. 2019 May 17;37(8):1030–1041. doi: 10.1002/stem.3026

Figure 2.

Figure 2

Insulin inhibits apoptosis during passaging. (A): Cell survival of dissociated cells 24 hours after plating on matrigel with or without insulin and rho‐associated protein kinase (ROCK) inhibitor (Y‐27632; ***, p < .001, n = 3). (B): Annexin V assay showing the percentage of apoptotic cells 4 hours after dissociation and plating on matrigel‐coated surface, with or without insulin or Y‐27632. (C): Flow cytometry analysis of Caspase 3/7 activity in dissociated embryonic stem (ES) cells 4 hours after plating on matrigel with or without insulin or Y‐27632 (Caspase 3/7, FITC‐A channel; FSC‐A, forward scattering). (D): Western blot showing the cleavage of Caspase 3 at Asp‐175 in cells cultured 4 hours on matrigel after dissociation with or without insulin and Y‐27632. Quantification is shown in Supporting Information Figure S2A. (E): Plots showing the cell survival 24 hours after plating on matrigel‐coated surface with or without the Caspase inhibitor Z‐VAD‐FMK, ROCK inhibitor Y27632, or insulin (***, p < .001, n = 3). (F): Cell proliferation of dissociated H1 ES cells during 72 hours after plating on matrigel‐coated surface comparing insulin effect to the Caspase inhibitor Z‐VAD‐FMK (*, p < .05; **, p < .01, n = 3; data are normalized to time zero cell count). Abbreviation: PI, propidium iodide (Annexin V, FITC‐A channel; PI, PE‐A channel).