Fig. 6: A developmental model system amenable to screening reveals the importance of EGF and FGF pathways in progenitor proliferation. | Nature Communications

Fig. 6: A developmental model system amenable to screening reveals the importance of EGF and FGF pathways in progenitor proliferation.

From: A 3D system to model human pancreas development and its reference single-cell transcriptome atlas identify signaling pathways required for progenitor expansion

Fig. 6: A developmental model system amenable to screening reveals the importance of EGF and FGF pathways in progenitor proliferation.

a PP-spheroids (from HUES4) were passaged into a 96-well plate at normal density and treatments in different conditions started after 2 days. After 8 days of treatment, ATP in each well was measured with a Cell titer-Glo assay, as a proxy for cell numbers. b Luminosity (arb. units) reflects the amount of ATP in each well. Results shown as mean ± SD (n = 3). c For validation, we performed EdU quantification by flow cytometry (PP-spheroids from HUES4 and SBAD3.4) or whole-mount immunostaining (fetal spheres) after 3 or 8 days of treatment. Treatment times were identical to the screening except for Infigratinib which was used for the last 24 h only. d EdU percentages for different conditions after 3, 8, or 1 (for Infigratinib) day of treatment, N = 3. Data shown as mean ± SEM. *P < 0.05. P values were determined by a two-sided Mann–Whitney test. e Representative Z-stack projections of whole-mount images showing PDX1, EdU, and DRAQ5 (nuclear stain) in PP-spheroids treated with different conditions. Images are representative of N = 4 validation immunostainings. Scale bar = 50 µm. See also Supplementary Fig. 5.

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