FIGURE 2

pkd2 knockdown in the DFCs rescues KV fluid flow pattern. (A) Immunostaining for cortical actin and rhodamine showing the result of a successful injection of pkd2 MO into DFCs, anterior (A’) and posterior (A”) panels in the different channels allow for better contrast/brightness balance; (B) Immunostaining for Pkd2 in WT and pkd2MO^DFCs embryos; (C) Representative images of KV architecture from one WT embryo and one pkd2 MO^DFCs injected embryo. (D) Quantification of the differences in length to width ratio in the KV of WT (n = 6) and pkd2 MO^DFCs (n = 9). (E) Cilia distribution in the antero-posterior axis of the KV of WT embryos (n = 22) and embryos injected with pkd2 MisMO (n = 6), pkd2 MO (n = 48) and pkd2 MO^DFCs (n = 15). (F) Quantification of the differences in cellular length, width, and height, in the same WT and pkd2 MO^DFCs injected embryos from panel (D). (G–I) Fluid flow heatmap and quantification of WT (n = 8), pkd2 MO^DFCs (n = 6) and pkd2 MO 1-cell stage embryos (n = 7), respectively. Asterisks represent statistical significance (Wilcoxon Test, p-value < 0.05). (J) 3D cilia length measurement in WT (n = 23), pkd2 MisMO (n = 8), pkd2 MO 1-cell stage (n = 25), rescue (n = 19), and pkd2 MO^DFCs (n = 10). (K) Motile/Immotile cilia ratio in live embryos injected with arl13b-mCherry mRNA (n = 8) and injected with pkd2 MisMO (n = 10), pkd2 MO (n = 6) and pkd2 MO rescued with Xenopus pkd2 mRNA (n = 5). Asterisks represent statistical significance (p < 0.05) with paired t-test. Scale bars 10 μm. L, left; R, right; A, anterior; P, posterior.