Validation of screen results that identify Ca2+ signaling as a regulator of DFC proliferation. (A) Diagram of SERCA function to pump Ca2+ from the cytoplasm into endoplasmic reticulum stores, and the action of the small molecules thapsigargin (Thaps) and cyclopiazonic acid (CPA) to inhibit SERCA. Created using BioRender.com. (B) Treating embryos with 1 μM Thaps or 100 μM CPA significantly reduced the mitotic index of DFCs as compared to negative control DMSO treated embryos. Bar graphs indicate average values and error bars represent one standard deviation. Each circle on the graphs represents results from an individual embryo. An unpaired two-tailed t-test with Welch’s correction was used for statistical analysis. * = significant difference. n = number of embryos analyzed. (C) Representative merged images of pHH3 staining in Tg(sox17:EGFP-caax) embryos treated with 1% DMSO (negative control), 1 μM Thaps, or 100 μM CPA. Arrows point out mitotic DFCs and asterisks point out mitotic neighboring (non-DFC) cells.
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