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. 2020 May 7;11:2241. doi: 10.1038/s41467-020-16049-3

Fig. 3. Screen to identify compounds that promote alpha cell identity.

Fig. 3

Small molecules targeting known pathways (43 compounds) were incubated with pre-alpha cells in quintuplicate for 96 h. a Schematic of screening approach. Primary screening results showing b the percentage of cells expressing both insulin and glucagon and c the percentage of cells expressing only glucagon. The data are presented as mean ± SEM (n = 5 biologically independent samples). The PKC activator PDBu significantly reduced the percentage of pre-alpha cells and increased the percentage of cells expressing glucagon. d Representative flow cytometry results of pre-alpha cells at stage 6 day 1, and stage 6 day 28 control (no treatment), or 28 days treatment with the PKC activator PDBu. e Immunofluorescence of clusters treated with and without PDBu for 28 days. Scale bar = 50 μm. f Additional PKC activators evaluated for their effect on pre-alpha cells (stage 6 day 1). All PKC activators increased the percentage of SC-alpha cells. The data are presented as mean ± SEM, significance calculated using an ordinary one-way ANOVA with Dunnett multiple comparison test (n = 3 biologically independent samples). g Stage 6 of protocol converts pre-alpha cells into SC-alpha cells.