12 research outputs found
Osbpl8 Deficiency in Mouse Causes an Elevation of High-Density Lipoproteins and Gender-Specific Alterations of Lipid Metabolism
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In vivo Functional selection identifies genes that enhance skeletal muscle regeneration after injury
Gene trap (GT) insertion site in Osbpl8.
<p>Insertion of the pGT vector in the 5<sup>th</sup> intron of the Osbpl8 gene in chromosome 10 generates a ÎČ-geo fusion mRNA transcript through the use of the En-2 splice acceptor (SA) contained in the vector. The insertion site (nt position indicated) was identified by PCR, using primers targeting gDNA in 5<sup>th</sup> intron of Osbpl8 gene (Fwd), and in the 1<sup>st</sup> intron of En2 (Rev1).</p
Liver tissue lipid content of the WT and Osbpl8KO mice.
*<p>T-test, comparison between genotypes, nâ=â6.</p
Non-cholesterol sterols in the plasma of WT and Osbpl8KO mice (nâ=â6).
#<p>T-test, comparison between genotypes; *p<0.05, **p<0.01.</p
Western blot analysis of HDL apolipoproteins in WT and KO mouse plasma and high-density lipoprotein fractions.
<p>A. Chow diet-fed animals; ApoA-II, ApoC-III, and ApoE (identified on the right) in the fasting plasma (Pla) of WT and KO mice of both genders (identified at the top); Bottom panel, ApoE in the HDL peak fractions of plasma fractioned on a Suprose HR 6 10/30 column. B. Western diet-fed animals; ApoE in the plasma (Pla) and HDL fractions (HDL) of WT and KO mice of both genders (identified at the top). The plasma loading was 3 ”l/lane of 1/40 dilution, and that of Superose 6 HR 10/30 HDL fractions 150 ”l/lane concentrated by acetone precipitation. The analysis was carried out for pooled plasma and HDL fractions isolated from the same pools; Chow diet, 3 animals/pool; Western diet, 6 animals/pool.</p
HDL catabolism in Osbpl8KO mice.
<p>Chow-fed WT and Osbpl8KO (nâ=â4â6) female (A) or male (B) animals were injected with Alexa568-labeled mouse HDL (40 ”g protein/animal), and the fluorescence signal (y-axis; mean ± s.e.m., % of starting value at 5 min post-injection) in their plasma as a function of time (x-axis) was measured as specified in Materials and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0058856#s4" target="_blank">Methods</a>.</p
Western blot analysis of ORP8 in KO mouse tissues and peritoneal macrophage.
<p>Total protein specimens (20 ”g protein/lane) of tissues with abundant ORP8 expression: spleen, brain, kidney, and liver (WT and Osbpl8KO animals), and of peritoneal macrophage (WT, Osbpl8+/â, and Osbpl8â/â) were analyzed by Western blotting with ORP8 and ÎČ-actin antibodies. ORP8 runs for an unknown reason as a doublet of bands with the apparent molecular masses of 97 and 101 kDa, as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0058856#pone.0058856-Yan1" target="_blank">[10]</a>.</p
Fasting plasma lipid concentrations of WT and Osbpl8KO mice before (nâ=â12) and after (nâ=â6) the diets.
#<p>TC, total cholesterol; FC, free cholesterol; TG, triglycerides; PL, choline-containing phospholipids; mmol/L.</p>##<p>T-test, comparison between genotypes; *p<0.05, **p<0.01, ***p<0.001.</p>§<p>apolipoprotein A-I; mg/ml.</p