Suzanne de Bruijn

96 Chapter 2 SUPPLEMENTARY FIGURES A B C s iRNA 1 s iRNA 2 s iRNA NT Percentage of ciliated cells siRNA 1 siRNA 2 siRNA NT 0 20 40 60 80 100 120 % of ciliated cells Cilium length siRNA 1 siRNA 2 siRNA NT 0 1 2 3 4 5 Cilium length ( µ m) Figure S1. Effect of KIAA1549 knockdown on ciliogenesis. hTERT-RPE1 cells were transfected with two different siRNAs targeting KIAA1549 (siRNA 1 and 2) and one non-targeting siRNA (siRNA NT). After transfection of the cells, ciliogenesis was induced and immunofluorescence was used for analysis. Primary cilia were stained using an anti-ARL13B antibody (red), and the transition zone was stained using an anti-RPGRIP1L antibody (green). (A) Percentage of ciliated cells calculated and (B) measured cilium lengths for hTERT-RPE1 cells transfected with the different siRNAs. (C) Images of ciliated cells transfected with one of the siRNAs. A close-up picture of the primary cilium is shown to visualize the transition zone. A B Long transcript (NM_001164665) siRNA 1 siRNA 2 siRNA NT 0.0 0.5 1.0 1.5 Relative expression to GUSB Short transcript (XM_935390) siRNA 1 siRNA 2 siRNA NT 0.0 0.5 1.0 1.5 Relative expression to GUSB *** ** ** Figure S2. Expression levels of KIAA1549 transcripts in HEK293T after knockdown using siRNAs. HEK293T cells were transfectedwith two different siRNAs targeting the long KIAA1549 transcript specifically (siRNA 1 and 2) and one non-targeting siRNA (siRNA NT) in two independent experiments. After transfection of the cells, expression levels of both the short and the long transcript were quantified using qPCR. (A) Using siRNA 1 and 2, a significant knockdown of the long KIAA1549 transcript was established. (B) Also, decreased expression levels of the short transcript were observed, which was found significant for siRNA 1. Significance was calculated using an unpaired t-test, ** p-value < 0.01, *** p-value < 0.001.