FIGURE

Fig. 3

ID

ZDB-FIG-120416-3

Publication

Ninov et al., 2012 - Different levels of Notch signaling regulate quiescence, renewal and differentiation in pancreatic endocrine progenitors

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Fig. 3

Notch signaling downregulation induces cell cycle progression of NRCs, whereas Notch signaling upregulation triggers their quiescence. (A-C) Larvae were treated with DMSO or 10 μM LY411575 from 4 to 6 dpf (52 hours) in the presence of EdU. (A) DMSO-treated control larva. Yellow arrowheads indicate EdU⁺ NRCs; white arrowheads indicate two endocrine cells along the IPD (projection of several planes). (B) The LY411575-treated larva showed an increase in the number of endocrine cells. Note that the majority of the new endocrine cells are EdU⁺ (arrows) and only two endocrine cells are EdU^– (arrowheads) (projection of several planes). Upon endocrine differentiation, however, NRCs exit the cell cycle (see supplementary material Fig. S3B,C). (C) Quantification of average percentage of EdU⁺ cells for A and B. Blue bar: on average, 43% of the NRCs in DMSO controls were EdU⁺ (140 cells counted in 7 larvae). Red bar: on average, 85% of the NRCs not undergoing endocrine differentiation in LY411575-treated larvae were EdU⁺ (140 cells counted in 7 larvae) (P<0.001). Green bar: the majority (65%) of new endocrine cells were EdU⁺ (129 cells counted in 7 larvae). Error bars indicate s.d. (D,E) Larvae were heat shocked at 4.5 dpf to induce mosaic expression of (D) ZdnSu(H)-myc or (E) myc-Notch1a-intra, and incubated in EdU for 24 hours (4.5 to 5.5 dpf). EdU incorporation was scored in cells positive or negative for the transgenes. (D) (ZdnSu(H)-myc)⁺ cells are identified by GFP expression (supplementary material Fig. S4A). Yellow arrowheads indicate five GFP⁺ NRCs, all of which are EdU⁺. Neighboring GFP^– NRCs (white arrowheads) incorporate EdU stochastically (projection of several planes). Eighty-one percent of GFP⁺ NRCs were EdU⁺ (104 cells counted in 52 larvae), whereas 42% of GFP^– NRCs were EDU⁺ (351 cells counted in nine larvae). (E) Myc staining marks myc-Notch1a-intra overexpressing cells (red) and Tg(Nkx2.2a:GFP) expression marks IPD cells (green). Myc⁺ IPD cells are EdU^– (yellow arrowheads). EdU incorporation is restricted to neighboring Myc^- IPD cells (white arrowheads) (projection of several planes). All images are lateral views, anterior towards the top, ventral towards the right. Scale bars: 20 μm.

Expression Data

Expression Detail

Antibody Labeling

Phenotype Data

Phenotype Detail

Acknowledgments

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