Cilia knockdown inhibits Notch signaling andklf2activation during ventricle regeneration. (A, B) Acetylated tubulin immunostaining in Tg(flk:GFP) heart showing cardiac cilia in control morphants (A) and ift88 morphants (B) at 3 dpf. Green, anti-GFP immunostaining; red, acetylated tubulin immunostaining. dpf, days post fertilization. (C) Quantification of cardiac cilia number in control morphants and ift88 morphants at 3 dpf (N = 15, 10 respectively). Mean + s.e.m. Student’s t-test, ****P < 0.0001. (D–G) Whole-mount in situ hybridizations indicated klf2a upregulation in control morphant ablated hearts (F) compared to control hearts (D) at 24 hpt, whereas this activation was blocked in ablated ift88 morphant hearts (G). (H–K) Whole-mount in situ hybridizations indicated klf2b upregulation in control morphant ablated hearts (J) compared to control hearts (H) at 24 hpt, whereas this activation was blocked in ablated ift88 morphant hearts (K). (L–O) Whole-mount in situ hybridizations indicated notch1b upregulation in control morphant ablated hearts (N) compared to control hearts (L) at 24 hpt, whereas this activation was blocked in ablated ift88 morphant hearts (O). (P–S) Confocal stack projections of ablated Tg(vmhc:mCherry-NTR; tp1:d2GFP) hearts indicated Notch signaling activation was inhibited in ift88 morphants at 12–24 hpt. Scale bars, 50 µm. hpt, hours post treatment. Dashed lines outline the hearts. Numbers indicate the ratio of representative staining observed
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