Figure 4.

Analysis of RECQL5-containing complexes by size exclusion column chromatography. For each fractionation experiment, 200 μl of sample was applied to a Superose 6 (10/300) column and serial elutants (0.5 ml each) were collected. The first 15 fractions were considered void volume and discarded, whereas individual subsequent fractions with an estimated mass larger than 130 kDa (the expected size of monomeric RECQL5) were saved and analyzed by Western blotting. A. Western blot results with fractions derived from the product of the affinity purification using antibodies specific for RECQLS. Note the detection of a prominent RECQL5 specific signal in fractions 18 and 19. B. Same as in A except that the input was the P.5 fraction, i.e., the input for the immunoaffinity purification experiment and the membrane was incubated sequentially with antibodies against RECQLS (top panel), RPB1 (middle panel), and BRG1 (bottom panel), respectively. Positions of three protein size markers (158, 669, and 2000 kDa, respectively) are shown.