Fig. 5: Knockdown of MIF affects the trafficking of KIF3a and IFT particles, including IFT20, IFT88 and IFT140.

a Representative image of RCTE cells transfected with MIF and control siRNAs for 48 h and serum starved for 24 h, then stained with KIF3a and Ac-αTub (red) antibodies and co-stained with DAPI (blue). Scale bars, 5 μm. b Stacked bar graph represented KIF3a distribution at primary cilia in RCTE cells transfected with MIF and control siRNAs. c Quantification of the intensity of KIF3a signals in RCTE cells transfected with MIF and control siRNAs. d Representative images of RCTE cells transfected with MIF and control siRNAs for 48 h and then serum starved for 24 h, and stained with IFT20 and Ac-αTub(red) antibodies and co-stained with DAPI (blue). Scale bars, 5 μm. e Stacked bar graph represented IFT20 distribution at primary cilia in RCTE cells transfected with MIF and control siRNAs. f Quantification of the intensity of IFT20 signals in RCTE cells transfected with MIF and control siRNAs. g Representative images of RCTE cells transfected with MIF and control siRNAs for 48 h and then serum starved for 24 h, and stained with IFT88 and Ac-αTub(red) antibodies and co-stained with DAPI (blue). Scale bars, 5 μm. h Stacked bar graph represented IFT88 distribution at primary cilia in RCTE cells transfected with MIF and control siRNAs. i Quantification of the intensity of IFT88 signals in RCTE cells transfected with MIF and control siRNAs. j Representative images of RCTE cells transfected with MIF and control siRNAs for 48 h and then serum starved for 24 h, and stained with IFT140 and Ac-αTub(red) antibodies and co-stained with DAPI (blue). Scale bars, 5 μm. k Stacked bar graph represented IFT140 distribution at primary cilia in RCTE cells transfected with MIF and control siRNAs. l Quantification of the intensity of IFT140 signals in RCTE cells transfected with MIF and control siRNAs. Numbers cells analyzed are indicated in the bars or above each dataset, for each group. Statistics were performed using one-sided one-way ANOVA followed by post hoc LSD test (b, e, h, k). Statistics were performed using student’s t-test (c, f, i, l). ***p < 0.001.