Oligomer distribution of recombinant adiponectin is altered in LH3^−/− knockout MEF cells.

<p>The oligomeric forms of recombinant adiponectin were separated with gel filtration chromatography and quantified by ELISA. Comparison of typical gel filtration elution profiles of recombinant adiponectin produced in (A) wild type and (B) LH3^−/− knockout MEFs indicates that HMW and MMW oligomers are not secreted from knockout cells. (C) Double transfection with adiponectin and LH3 constructs normalized the elution profile in LH3^−/− knockout MEFs. (D) Recombinant adiponectin produced in LH mutant MEFs formed similar oligomers as the wild type. The adiponectin oligomers corresponding to the peaks seen with gel filtration chromatography were also detected on immunoblot (E) separated under non-reducing and non-heat-denaturing conditions. Equal volumes of concentrated cell culture media were used in all analysis. Representative elution profiles are shown. Abbreviations: WT = wild type; KO = LH3^−/− knockout; LH3 = lysyl hydroxylase isoform 3; MUT = LH mutant.