Fig. 4.

Reduced [1,2-¹⁴C]acetate incorporation in the polar lipids PIM₆, LM and LAM from M. marinum ΔMMAR_2380. (a) Extracted crude lipoglycans from acetate-labelled delipidated cells (25 μg) from M. marinum wild-type, ΔMMAR_2380 and complemented ΔMMAR_2380 (ΔMMAR_2380 comp.) were counted for the incorporation of [1,2-¹⁴C]acetate and analysed by SDS-PAGE/autoradiography. Shown is the average of two independent experiments. (b) [1,2-¹⁴C]Acetate-labelled M. marinum cultures were processed and polar lipids were applied (25 000 c.p.m.) to the corners of 6.6×6.6 cm pieces of aluminium-backed TLC plates and analysed using the 2D solvent system E as described in Methods. Plates were dried and autoradiograms were produced by overnight exposure of Kodak X-Omat AR film to the TLC plates to reveal [1,2-¹⁴C]acetate-labelled lipids. PI, non-mannosylated, diacylated phosphatidylinositol anchor; PIM₂, phosphatidylinositol dimannoside; PIM₆, phosphatidylinositol hexamannoside; Ac₁PIM, tri-acylated PIM; Ac₂PIM, tetra-acylated PIM. Unassigned spots in the figure are lipooligosaccharides, and in the upper-right corner are diphosphatidylglycerol, phosphatidylethanolamine and unknown phospholipids (Burguière et al., 2005). Two independently prepared lipid extracts per strain were tested.