<p><b>A</b>, schematic depicting the pull-down assay on a folate affinity column. <b>B</b>, endogenous ALDH1L1 interacts strongly with immobilized folinic acid (after loading NIH3T3 cell lysate on the affinity column, the following eluted fractions were collected and analyzed by SDS-PAGE/Western blot assay with ALDH1L1-specific antibody: <i>lane 1</i>, washing buffer; <i>lane 2</i>, 0.5 M KCl; <i>lane 3</i>, 1.0 M KCl; <i>lane 4</i>, 2.0 KCl; <i>lane 5</i>, 5 mM folic acid in 2.0 KCl; <i>lane 6</i>, 20 mM folic acid in 2.0 M KCl; <i>lane 7</i>, purified ALDH1L1 standard; <i>St</i>, molecular masses standards (indicated by numbers in kDa on the <i>left</i>). <b>C</b>, samples as in <b>B</b> probed with CHIP-specific antibody. <b>D</b>, samples as in <i>B</i> probed with HSP90-specific antibody. <b>E</b>, Immunoprecipitation of NIH3T3 cell lysate with ALDH1L1-specific (<i>left panels</i>) or HSP90-specific (<i>right panels</i>) antibody followed by SDS-PAGE/Western blot assay with ALDH1L1-specific, HSP90-specific and CHIP-specific antibodies. In each experiment (ALDH1L1 or HSP90 pull-down) the same blot was stripped twice and reprobed. Numbers indicate molecular masses (kDa) for standards.</p
