Fig. 3: IFT74-related Joubert syndrome (JBTS) is associated with defects of cilia.
From: Disrupted intraflagellar transport due to IFT74 variants causes Joubert syndrome
(a) Representative images of control and patient fibroblasts stained for acetylated tubulin (green), CEP164 (red), and DAPI (blue). Cells were treated with serum starvation for 48 hours before fixation. Scale bars, 30 μm. (b) Quantification of cells with cilia from healthy controls and JBTS patients. (c) Quantification of ciliary length of the fibroblasts. (d) Quantification of cells with cilia after the treatment of serum starvation for 0 hours, 5 hours, and 12 hours. (e) Western blots of whole cells probed with the indicated antibodies. (f) Quantification of IFT74 protein levels relative to the β-actin loading control. Statistical significance was determined by the t-test (**p < 0.01). (g) Western blots of protein levels of RPE1 cells by probing indicated antibodies. Wild-type cells or cells stably expressing wt-IFT74-Flag or Q179E-IFT74-Flag were transfected with control small interfering RNA (siRNA) or siRNA targeted IFT74. (h–j) Representative images of fibroblasts stained for IFT74, IFT88, or IFT140 and costained for ARL13B to indicate cilia. Scale bars, 3 μm. (k–m) Quantification of IFT protein levels (in h–j) in cilia. (n–p) Representative images of fibroblasts stained for TMEM67 (red) and ARL13B (green) (n), TCTN1 (red) and ARL13B (green) (o), OFD1 (red) and ARL13B (green) (p). Scale bars, 1.5 μm. Error bars represent SD.
