adora2ab depletion induces cystogenesis and ciliary defects in zebrafish embryos. (a) Dose-dependent increase in cyst formation following adora2ab TBM injection in wild-type embryos. (b) Dose-dependent increase in cyst formation following adora2ab SBM injection. (c) Quantification of cyst formation in rescue experiments. The co-injection of adora2ab mRNA (30 ng/μl) significantly reduces cyst formation in adora2ab TBM-injected embryos. The control morpholino shows minimal cyst formation. (d) Confocal microscopy analysis of pronephric cilia at 48 hpf using acetylated tubulin immunostaining (magenta) in Tg(wt1b:GFP; cldnb-ly:GFP) embryos. adora2ab TBM injection results in an irregular cilia morphology, while SBM-injected embryos maintain a normal ciliary structure. Bar = 10 µm. (e) The sgRNA-mediated targeting of adora2ab leads to increased cyst formation in wild-type embryos. (f) Enhanced cyst formation in nphp1/nphp4 morphants co-injected with adora2ab TBM. (g) Analysis of cyst formation in nphp1ex15-del4;nphp4sa38686 homozygous mutants and wild-type siblings following adora2ab sgRNA injection (n = 226 and 235 embryos). Each circle represents an independent experiment. Data are presented as mean ± SEM.
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