1. DHA induces peroxisome elongation and division in a Pex11pβ-dependent manner.
DHA induces peroxisome elongation and division in a Pex11pβ-dependent manner. (A) AOx-deficient fibroblasts were treated with a control dsRNA (upper row) or PEX11β #2 dsRNA (lower row) for 48 hours. Cells were cultured for an additional 12 hours (Ab,Ae) and 24 hours (Aa,Ac,Ad,Af) in the absence (Aa,Ad) or presence (Ab,Ac,Ae,Af) of 150 μM DHA and then stained with an anti-Pex14p antibody. Insets are higher-magnification images of the boxed regions. Scale bars: 10 μm and 2 μm (insets). (B) AOx-deficient fibroblasts were treated with control dsRNA or two different test dsRNAs (PEX11β #1 and PEX11β #2) for 48 hours. Pex11pβ levels were verified by immunoblot analysis using an antibody to Pex11pβ. Actin was used as a loading control. (C) AOx-deficient fibroblasts were treated as in A with control, PEX11β #1, PEX11β #2 or PEX11β #1 plus PEX11α #1 dsRNAs. Cells were cultured for an additional 24 hours in the absence or presence of 150 μM DHA. Peroxisome abundance per cell was determined as in Fig. 2B. Data represent the means ± s.d. of three independent experiments; *P<0.01. (D) AOx-deficient, control, and PEX10-deficient fibroblasts were cultured for 20 hours in the absence (left panels) or presence (right panels) of 150 μM DHA and then treated for 30 minutes with 1 mM DSP. Cell lysates were fractionated by sucrose density gradient ultracentrifugation and each fraction analyzed by immunoblot using antibodies to Pex11pβ, DLP1 and Fis1. Upward arrowhead indicates the higher molecular-mass oligomer of Pex11pβ.
Akinori Itoyama et al. J Cell Sci 2012;125:
© 2012.