Functional role of DLPI in peroxisome proliferation.

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Functional role of DLPI in peroxisome proliferation. Functional role of DLPI in peroxisome proliferation. (A) AOx-deficient fibroblasts were cultured for 16 hours in the absence (Aa,Ab) or presence (Ac,Ad) of 150 μM DHA and then treated for an additional 8 hours with DMSO (Aa,Ac) or 40 μM dynasore (Ab,Ad). Cells were stained with an anti-Pex14p antibody. (B) Peroxisome abundance per cell was determined as in Fig. 2B. (C) AOx-deficient fibroblasts were treated for 48 hours with a control dsRNA (left panel) or DLP1 #2 dsRNA (right panel). Cells were cultured for an additional 24 hours in the absence (Ca,Cb) or presence (Cc,Cd) of 150 μM DHA and then stained with an anti-Pex14p antibody. (D) AOx-deficient fibroblasts were treated for 48 hours with a control dsRNA or two different test dsRNAs (DLP1 #1 and DLP1 #2). DLP1 levels were verified by immunoblot analysis using an anti-DLP antibody. Actin was used as a loading control. (E) Peroxisome abundance per cell was determined as in Fig. 2B. (F) Control (Fa–Fc) and AOx-deficient fibroblasts (Fd–Fi) were cultured for 24 hours in the absence (Fa–Ff) or presence (Fg–Fi) of 150 μM DHA. Cells were stained with antibodies to Pex14p (Fa,Fd,Fg) and DLP1 (Fb,Fe,Fh); the merged view of the two different proteins is shown in Fc, Ff, and Fi. Insets are higher-magnification images of the boxed regions. Scale bars: 10 μm and 2 μm (insets). Data represent the means ± s.d. of three independent experiments; *P<0.01. Akinori Itoyama et al. J Cell Sci 2012;125:589-602 © 2012.