Ofd1 expression levels in cells affect cilia number and length.

<p>(A) shRNA-mediated (pSUPER-EGFP1-shRNA-Ofd1) down-regulation of Ofd1 in R28 cells determined using qRT-PCR and Western blotting analysis. Knockdown efficiency was 52%. *P<0.05. (B–C) Ciliated cell number and cilia length decreased after shRNA mediated down-regulation of Ofd1 in R28 cells (shRNA), compared with pSUPER-scramble shRNA (without EGFP) treated cells (control). *P<0.05. Unit: um. For quantification results, three replicates were performed but only one band for three representative samples per group was shown. (D) Representative image of Acetylated-α-tubulin immunostaining results. Acetylated-α-tubulin-cy3, marker of cilia axoneme, was used to demonstrate cilia in R28 cells. Cells were transfected with pSUPER-EGFP1-shRNA-Ofd1 for 48 h (shRNA), or pSUPER-scramble shRNA construct (without EGFP) as control. Magnification 600× under microscopy. Scale bar: 50μm. Enlarged: the magnified image of representative part in white box. White arrow: Acetylated-α-tubulin-cy3 staining presented cilia.