Figure 4.—

Immunodetection of C. reinhardtii CrSUMO96, CrSUMO148, and CrSUMO-like89A with anti-CrSUMOs and anti-AtSUMO-1 antibodies. Overexpression and purification of 6× His-tagged CrSUMOs from E. coli, antisera production, and immunoblot detection are shown. CrSUMO96 overexpressed in E. coli and affinity purified on a 6× His column (lanes 2, 5, 8, 11, 14, and 17), overexpressed and purified CrSUMO148 (lanes 3, 6, 9, 12, 15, and 18), and overexpressed and purified CrSUMO-like89A (lanes 4, 7, 10, 13, 16 and 19) were separated on 12% SDS–PAGE and stained with Coomassie blue (lanes 1–4) or detected after immunobloting using anti-6× His antibody (lanes 5–7), anti-AtSUMO1 antiserum (lanes 8–10), anti-CrSUMO96 antiserum (lanes 11–13), anti-CrSUMO148 antiserum (lanes 14–16), and anti-CrSUMO-like89A antiserum (lanes 17–19).