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Fig. 7
INPP5B can partly rescue the defect of OCRL in primary cilia development.
(A) INPP5B protein levels in HEK293T cells. GFP-Inpp5b lentivirus was generated in HEK293T cells; both GFP-Inpp5b and endogenous INPP5B were immunoblotted in 40 microgram lysates; beta-actin levels are shown. (B) NHF 558, Lowe 1676, and Lowe 3265 fibroblasts were transduced with control, Inpp5b or Inpp5b ΔCAAX lentivirus, serum-starved for 48-hours, and immunostained with acetylated alpha -tubulin. Quantification of cilia length is shown (n >100 cilia, three independent experiments, unpaired t-test, * p = 0.004, ** p = 0.006). (C) Inpp5b mRNA partly rescued the loss of Ocrl. KV cilia of zebrafish embryos injected with ocrl MO (4 ng) with Inpp5b WT mRNA (500 ng) and ocrl MO (4 ng) with Inpp5bΔCAAX mRNA (500 ng) at 6-somite stage were immunostained with acetylated α-tubulin (red), representative images are shown (dash line indicates border of KV). Scale bar 10 micron. (D) Quantification of length of KV cilia in zebrafish embryos injected with ocrl MO (4 ng) with Inpp5b WT mRNA (500 ng) and ocrl MO (4 ng) with Inpp5bΔCAAX mRNA (500 ng). (N >20 embryos, three independent experiments, unpaired t-test, * p = 2.2E-04).