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Fig. 4. Ciliary defects in Xenopus cfap43 morphant and crispant larval skin.
(A) (a) Kymographs of ciliary motility of single MCCs were generated from control (co) wt, TBMO-, sgRNA1-and sgRNA2-injected specimens. (b) Statistical evaluation of results from 3 independent experiments with 15 embryos each and 5 analyzed MCCs per embryo revealed elevated ciliary beat frequencies upon cfap43 loss-of-function. (B) (a) Schematic depiction of the region used for flow analysis in control wt and manipulated stage 32 embryos. Embryo scheme taken from https://www.xenbase.org/anatomy/alldev.do. (b) Maximum intensity projections of single control wt, TBMO-, sgRNA1-and sgRNA2-injected specimens represent cilia-generated flow. (c) Evaluation of bead transport from 3 independent experiments with 8 analyzed specimens each shows reduced mean velocities in cfap43 morphants and crispants. (C) cfap43 mRNA transcripts were reduced in cfap43 crispant embryos (b, d) at stage 33, with a focus on nephrostomes (a, b) and MCCs (c, d). Morphant Embryos were injected with 0.5 pmol TBMO. (D) IF staining of basal bodies (centrin GFP) and basal feet (Tubg1) in skin MCCs of stage 32 wildtype and crispant specimens. Scale bars: C = 100 μm, D = 10 μm ***, p<0.001. |
