SERCA inhibitor treatments during epiboly reduce the number of ciliated KV cells and disrupt left-right patterning of the embryo. (A) Representative images of the ciliated KV in Tg(sox17:EGFP-caax) embryos treated with 1% DMSO (control) or 1 μM Thapsigargin (Thaps). KV cells are labeled with membrane-localized EGFP expression (green), and KV cilia are labeled with anti-acetylated tubulin antibodies (magenta). (B–D) Treating embryos with 1 μM Thapsigargin during epiboly stages reduced the area of KV (B), the number of cilia in KV (C), and the number of cells in KV (D) relative to DMSO treated controls. 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. (E) Representative images of normal left-sided spaw expression (arrows) in a DMSO control embryo and bilateral expression in a thapsigargin treated embryo. (F) Most DMSO control embryos had left-sided spaw expression, whereas spaw was largely bilateral, and in some cases absent or right-sided in thapsigargin treated embryos. (G) Representative images of normal leftward heart jogging (dashed arrow) in a DMSO treated control embryo and midline jogging in a thapsigargin treated embryo. The heart tube was labeled by EGFP expression in cardiomyocytes. (H) Leftward heart jogging was observed in most control embryos. In contrast, the heart often jogged along the midline or to the right in thapsigargin treated embryos. n = number of embryos analyzed.
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