Fig. 2
Depletion of MPP9 leads to aberrant ciliogenesis. a Immunostaining of acetylated-tubulin (Acet-Tub, red) and MPP9 (green) in normal and serum-free medium-treated hTERT RPE-1 cells. b Quantification of the percentage of cells with MPP9 at distal ends of the mother centrioles after serum starvation. c Immunoblots showing depletion of MPP9 by siRNAs (#1 and #2) transfection and rescue of MPP9 expression by overexpressing Flag-tagged siRNA-resistant MPP9 (Flag-ResMPP9). Tubulin was used as a loading control. Relative amounts of MPP9 were quantified and normalized to tubulin. d Quantification of ciliogenesis in control-siRNA, MPP9-siRNA, or CP110-siRNA, or MPP9-siRNA together with Flag-ResMPP9-rescued hTERT RPE-1 cells. e Immunostaining of Centrin-3 (red) and Arl13B (green) in MPP9-siRNA-treated and Flag-ResMPP9-rescued hTERT RPE-1 cells. DNA was stained with DAPI (blue). f Whole mpp9+/+ and mpp9–/– embryos at embryonic day (E) 10.5. g The male mpp9+/+ (left) and mpp9–/– (right) mice at 1 month after birth. h Line graph showing the body weight of mpp9+/+ and mpp9–/– mice at the indicated times (n = 6 mice for each group). i Immunoblots of MPP9 and CEP97 in the kidneys of mpp9+/+ or mpp9–/– mice at 1 month after birth. GAPDH was used as a loading control. j Kidneys from mpp9 +/+ and mpp9 –/– mice (n = 3 mice for each group) at 1 month were stained with Acet-Tub (red) and DAPI (DNA; blue). White dashed lines represent the border of each renal tubule. k The percentage of ciliated cells from j. For b, d, h, and k, bars represent the means ± S.E.M for three independent experiments. n.s., not significant, *p < 0.05, **p < 0.01, ***p < 0.001, as determined by unpaired two-tailed Student’s t-test (b, h, and k), one-way ANOVA analysis (d). Scale bars: 1 mm (f); 5 μm (e main, j); 1 μm (a)
