Figure S7.

The functional impairment of PRPF8 in pre-mRNA splicing does not affect ciliogenesis (related to Fig. 7). (A) GFP-vector (−), GFP-PRPF8 WT, or ΔMPN mutant were transfected into HEK293T cells. The cell lysates were pulled down by GST-Ub4, and then GFP proteins were detected by anti-GFP antibody. GST-Ub4 was stained by Ponceau S. (B) RPE-1 cell lysates in Fig. 7 D were analyzed by immunoblotting with the indicated antibodies. (C) GFP-vector (−), GFP-PRPF8 WT, or GFP-PRPF8 R2310K mutant were transfected into control or PRPF8-depleted RPE-1 cells and serum starved for 48 h. The cells were stained with the indicated antibodies. Scale bars, 5 µm (main image) and 1 µm (magnified region). (D) Quantification of the ciliated cells in C. Data are presented as mean ± SD of three independent experiments. A one-way ANOVA test was performed followed by Bonferroni’s multiple comparisons. ***, P < 0.001 by one-way ANOVA with Bonferroni’s multiple comparisons test. IB, immunoblot; tub, tubulin.