HeLa cells were lysed in the presence of 0.1% Triton X-100 and the 10,000×g soluble supernatants were applied to sepharose CL-6B gel filtration columns as described in Materials and methods. GibertBenjamin EckelBénédicte FasquelleLydie MoulinMaryline BouhallierFrantz GoninVincent MellierGregory SimonStéphanie Kretz-RemyCarole ArrigoAndré-Patrick Diaz-LatoudChantal 2012 <p>29, 66, 150, 200, 443, 669 and 2000 (kDa) indicate the apparent native size of gel filtration markers. <i>A</i>, The presence of Hsp27, HDAC6, STAT2 and procaspase-3 in pooled fractions eluted from the columns was detected by western blot analysis. <i>B</i>, interactions between Hsp27 and client proteins analyzed by Co-IP experiments using a goat polyclonal anti-Hsp27 antibody. Column fractions corresponding to either procaspase-3, STAT2 or HDAC6 were pooled and used for immunoprecipitation. Immunoprecipitated proteins and input cell lysates were analyzed side by side in immunoblots probed with the indicated antibodies. A representative negative control of Co-IP was performed with a control antibody and revealed (here the anti procaspase 3).</p>