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. 2024 Sep 30;28(3):109–119. doi: 10.12717/DR.2024.28.3.109

Fig. 3. Loss of Dstn results in length reduction of primary cilia in Xenopus and hRPE1 cells.

Fig. 3.

(A) Acetylated tubulin immunostaining of dstn-deficient embryos revealed a considerable reduction in the length of neural tube primary cilia of Xenopus compared with control embryos (scale bar=10 μm). Statistical analysis showed that the dstn morphants exhibited a length of primary cilia less than 5 μm compared with approximately 8 μm long cilia in control embryos. (B) RT-PCR analysis revealed the specificity of DSTN siRNAs as the expression of DSTN was significantly reduced in RPE1 cells transfected with DSTN siRNAs. GADPH was used as the internal control. (C) Western blotting showed that the protein expression of DSTN was considerably reduced in DSTN siRNAs transfected hRPE1 cells. β-actin was used as the loading control. (D) hRPE1 cells were transfected with DSTN siRNAs and serum-starved for 48 hrs to induce ciliogenesis. As analyzed by immunostaining with acetylated tubulin, depletion of DSTN induced shorter primary cilia in hRPE1 cells with less than 2.5 μm compared with 5 μm long primary cilia in control hRPE1 cells (scale bar=10 μm). Statistical analysis of data revealed that the length of primary cilia was significantly reduced in DSTN-depleted cells as compared to the control cells. * p<0.05, ** p<0.01, *** p<0.001, **** p<0.0001.