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. 2022 Oct 12;11:e80165. doi: 10.7554/eLife.80165

Figure 2. Differentiation of CEP83−/− human-induced pluripotent stem cells (hiPSCs) to intermediate mesoderm cells (day 7) is associated with defective ciliogenesis.

(A) The schematic illustrates the applied differentiation protocol of hiPSCs, as previously described by Takasato et al., 2015. (B–C) WT and CEP83−/− cells on 7 days of culture (D7) of differentiation did not show overt morphological differences by brighfield microscopy. (D–E) Representative images of WT and CEP83−/− cells on D7, immunostained for acetylated tubulin (green) and nuclei (DAPI, blue), revealing fewer and elongated cilia in CEP83−/− cells. (F) Quantitative analysis of the percentage of ciliated cells in WT and CEP83−/− cells (D7). (G) Quantitative analysis of the ciliary length in WT and CEP83−/− cells (D7). n=3 clones per group. ****p<0.0001. Bar = 50 μm. See Figure 2—figure supplement 1.

Figure 2.

Figure 2—figure supplement 1. Loss of CEP83 in organoids results in defective ciliogenesis.

Figure 2—figure supplement 1.

(A) Immunofluorescence staining of wildtype (WT) and CEP83−/− organoids for acetylated tubulin (green), CEP83 protein (red), and nuclear staining 4',6-diamidino-2-phenylindole dihydrochloride (DAPI). Note CEP83 localization at the base of the cilium in WT organoids. (B) Quantitative analysis of ciliated cells showing downregulation of the number of ciliated cells in CEP83−/− organoids, associated with longer cilium formation (C). n=3 clones per group. Data are mean ± SD. ****p<0.0001. Panel A–B: Bar = 50 μm.