Fig. 7: Ceramide, pPKCζ and CDC42 colocalize with the ciliary marker Arl13b in primary cilia and deletion of IFT20 impairs ceramide ciliary distribution and β-catenin apical localization in vivo. | Communications Biology

Fig. 7: Ceramide, pPKCζ and CDC42 colocalize with the ciliary marker Arl13b in primary cilia and deletion of IFT20 impairs ceramide ciliary distribution and β-catenin apical localization in vivo.

From: Primary cilia control cell alignment and patterning in bone development via ceramide-PKCζ-β-catenin signaling

Fig. 7

Immunofluorescence staining of paraffin embedded tissue sections prepared from femur and tibia was performed to detect the Ceramide, pPKCζ and CDC42 using mouse-derived monoclonal antibodies in combination with ciliary marker, rabbit-derived polyclonal Arl13b antibody. Micrographs representing Z-stacked 3D-deconvolution processed images captured from the trabecular bone regions of postnatal day 11 IFT20f/f femur using Leica DMI6000 inverted epifluorescence microscope under ×100 lens. Colocalization of Ceramide (a–c), phosphorylated PKCζ (d–f) and CDC42 (g–i) with Arl13b were detected. Similar experiment was also performed using tamoxifen injected IFT20f/f mouse (j, l, n) and IFT20f/f Col1-creERT (k, m, o). The ciliary distribution of Ceramide was abolished as the Arl13b pattern was lost in the IFT20 deficient cells using tamoxifen injected IFT20f/f; Col1-creERT mouse (k, m, o). Scale bar (a–m) 10 μm, scale bar (n, o) 5 μm.

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