Fig. 5

Modulating Cftr activity restores lumen size in myo1d MZ mutant embryos. a, b Knockdown of myo1d in TgBAC(Cftr-GFP) embryos did not affect CFTR apical GFP localization (n = 6 control MO, n = 6 myo1d MO). c–e Activation of Cftr function in the absence of myo1d MZ mutant expanded lumen area. Representative Z-stack image showing that Forskolin and IBMX (FIBMX) increased KV lumen in WT and myo1d MZ embryos (c). Treating with ouabain, an inhibitor of Na⁺/K⁺ ATPase decreased the KV lumen area in WT and myo1d mutant embryos. Numeration of lumen area with treatments are shown in (WT-DMSO: n = 8, WT-ouabain: n = 10, WT-FIBMX: n = 4, myo1d MZ-DMSO): n = 19, myo1d MZ-ouabain: n = 7, myo1d MZ-FIBMX: n = 12) (d). Cardiac looping defects were increased in WT embryos treated with FIBMX or ouabain but did not increase in myo1d MZ mutants (e). Unpaired t test and one-way ANOVA were conducted. *p < 0.05, ***p < 0.001, ****p < 0.0001 represents statistical significance. ns not significant. Data shown are mean ± SEM. L lumen, scale bar = 15 µm (a), 50 µm (c)